Gibco™ 24 Hours of Stem Cells™ virtual event—2016 Speakers
Silke Rickert-Sperling, PhD
Charité Medical Faculty and the Max Delbrück Center for Molecular Medicine
Charité – Universitätsmedizin Berlin
Genetic variability of ps-iPSC and related blood and fibroblasts—somatic mutations
The generation of induced pluripotent stem cells (iPSCs) from adult easily accessible somatic tissues was introduced ten years ago. This technology has revolutionized our opportunities to study human disorders including complex genetics and develop novel therapeutic concepts for regenerative medicine. By differentiating iPSCs to different cell types and stages, we can gain insights into developmental and disease driving molecular networks. Yet, a discussed question of the iPSC model system is the extent to which they retain residual phenotypes from their precursor, which impacts on the study of transcription networks. Moreover, it is open to which degree somatic mutations impact on functional studies of human genetic disorders when patient-specific iPSCs are applied. Here, we will tackle this question in the course of studying healthy and diseased siblings with Tetralogy of Fallot, a frequent congenital heart defect of multigenic origin. Routine genetic screenings are mainly based on genetic material derived from either whole blood or cheek swap sampling. ps-iPSCs are typically derived from dermal fibroblast (skin biopsies) or whole blood derived immortalized lymphoplastoid cell lines (LCLs). We will discuss whole genome frequency and impact of somatic acquired mutations on gene expression and functional studies of complex genetic disorders using ps-iPSCs.
Prof. Dr. Silke Rickert-Sperling holds a doctoral degree in cardiac physiology and studied medicine from 1991 up to her full medical license in 1997. During her education she went from Berlin to New York, San Diego, Chicago, and Rochester. Afterwards she worked as a MD at the German Heart Center of Berlin. In 1999, she began her postdoctoral research at the department of Prof. Dr. Lehrach at the Max Planck Institute for molecular genetics and there she became head of the research group “Cardiovascular Genetics” in 2001. She holds a habilitation in molecular biology and bioinformatics. In 2011, she was honored with a Heisenberg-Professorship and became full-professor for Cardiovascular Genetics at the Medical Faculty of the Charité in Berlin. She is coopted professor at the Faculty of Biology, Chemistry and Pharmacy of the Freie Universität Berlin. She leads an interdisciplinary research group (molecular biology and bioinformatics) at the ECRC. As guest scientist, she continues to collaborate with the Max Planck Institute for molecular genetics. Her research activities focus on understanding the molecular basis of human cardiovascular disorders in particular congenital heart malformations and cardiac dysfunction. Using systems biology approaches, she studies cardiac (dys)development and muscle maturation in human and mice. She investigates a broader range of disease-associated genes and epigenetic modulators in the genomic, transcriptomic and proteomic context. She hopes to combine knowledge of molecular etiologies and mechanisms to eventually improve preventive and therapeutic opportunities for patients. She is principal investigator and co-coordinator of various European efforts (HeartRepair, CardioGeNet, and CardioNeT) and principal investigator of the Berlin Institute of Health (BIH). She was board member of the European Society of Human Genetics and is Fellow of the European Society of Cardiology (ESC) as well as board member of the ESC Working Group on Development, Anatomy and Pathology.
David Schaffer, PhD
Professor of Chemical and Bimolecular Engineering, Bioengineering, and Neuroscience
Director of the Berkeley Stem Cell Center
University of California, Berkeley
Molecular Elucidation and Engineering of Stem Cell Fate Decisions
Stem cells play critical roles in the development of organisms, as well as in the maintenance and repair of organs and tissues throughout adulthood. Advancing our understanding of mechanisms that control stem cell behavior – in particular their two hallmark properties of self-renewal and differentiation into specialized cells – will enable these cells to be increasingly harnessed to repair tissues damaged by disease or injury. Stem cells reside within specialized microenvironments or niches that present them with a spectrum of regulatory signals to control their behavior. In particular, the niche presents stem cells with a range of molecular cues, and it has also been become increasingly apparent that key biophysical features of the environment modulate the presentation of this biochemical information. For example, spatial and temporal variation in the presentation of cues is important information that can impact fate decisions and tissue structure. In addition, the tissue matrix can have variable bulk mechanical properties and surface topographical properties depending on how it’s assembled.
We have created several technology platforms to investigate these problems, and in particular to understand and control the differentiation of adult neural stem cells and human pluripotent stem cells into neurons. First, we are developing and harnessing optogenetics as a system to investigate how cellular signaling dynamics impact fate decisions. Second, we develop bioactive, synthetic material systems to investigate the effects of cell-matrix and cell-cell interactions on cellular function. Finally, we work towards translating the basic information that emerges from both of these efforts into safe, scalable, fully defined, robust culture and implantation systems for stem cell based regenerative medicine efforts to treat human disease.
David Schaffer is a Professor of Chemical and Biomolecular Engineering, Bioengineering, and Neuroscience at University of California, Berkeley, where he also serves as the Director of the Berkeley Stem Cell Center. He received a B.S. in Chemical Engineering from Stanford University in 1993 and a Ph.D. in Chemical Engineering from the Massachusetts Institute of Technology in 1998. He then conducted a postdoctoral fellowship at the Salk Institute for Biological Studies in La Jolla, CA before becoming a faculty member at the University of California at Berkeley in 1999. At Berkeley, Dr. Schaffer applies engineering principles to enhance stem cell and gene therapy approaches for neuroregeneration, work that includes novel approaches for molecular engineering and evolution of new viral vectors as well as new technologies to investigate and control stem cell fate decisions. David Schaffer has received an NSF CAREER Award, Office of Naval Research Young Investigator Award, Whitaker Foundation Young Investigator Award, and was named a Technology Review Top 100 Innovator. He was also awarded the American Chemical Society Marvin Johnson Award in 2016, the American Chemical Society BIOT Division Young Investigator Award in 2006, the Biomedical Engineering Society Rita Shaffer Young Investigator Award in 2000, and was inducted into the College of Fellows of the American Institute of Medical and Biological Engineering in 2010.
Budd Tucker, PhD
Stephen A. Wynn Professor in Regenerative Ophthalmology
Associate Professor of Ophthalmology and Visual Sciences
University of Iowa Carver College of Medicine
Using Patient-derived iPSCs to model and treat inherited retinal degenerative blindness
Inherited retinal degenerative disorders such as retinitis pigmentosa are characterized by death of the light sensing photoreceptive neurons of the outer retina. Like the rest of the CNS, the retina has little capacity for endogenous regeneration, and as a result, photoreceptor cell death causes debilitating irreversible blindness. Gene augmentation has the potential to prevent photoreceptor cell death, while cell replacement could actually repopulate the retina with new functioning photoreceptor cells and restore vision. In this talk I will show how we are using patient-specific iPSCs to evaluate disease pathophysiology, test novel gene-based therapeutics and develop autologous photoreceptor cell replacement for the treatment of retinal degenerative blindness.
Dr. Tucker was born and raised in a small fishing village (population <100) on the northern tip of Newfoundland, Canada. He attended Sir Wilfred Grenfell College in Corner Brook Newfoundland, where in 2001 as an undergraduate student he received his bachelors degree in Psychology. In 2006 Dr. Tucker went on to complete his Ph.D. degree in neuroscience at Memorial University of Newfoundland's School of Medicine. He subsequently completed a 3 year post-doctoral fellowship at the Schepens Eye Research Institute, Harvard Medical School, where in 2009 under the Mentorship of Dr. Michael J. Young was promoted to the rank of faculty. In 2010 Dr. Tucker joined the Department of Ophthalmology and Visual Science at the University of Iowa where he is currently an Associate Professor of Ophthalmology and Visual Science. Dr. Tucker has a long-standing interest in the treatment of inherited retinal degenerative diseases such as retinitis pigmentosa (RP), Stargardt disease, Usher Syndrome and age-related macular degeneration (AMD). His lab is focused on combining state-of-the-art patient-specific stem cell, gene augmentation/genome editing and tissue engineering based technologies to develop treatments for inherited retinal degenerative blindness.
PANEL DISCUSSION: From bench to BLA – a review of common regulatory questions
Joyce Frey- Vasconcells, PhD
Cell and Gene Therapy Regulatory Consultant
Frey-Vasconcells Conculting, LLC
Michael Mendicino, PhD
Owner, Chief Consultant & Advisor
Hybrid Concepts International
Kelli Tanzella, PhD
Senior Director, Global Regulatory Affairs, Clinical & Compliance
Thermo Fisher Scientific
From bench to BLA - a review of common regulatory questions
This panel will discuss quality requirements of ancillary materials used in cell therapy manufacturing, relevant standards and how requirements vary by phase. Participants will learn what to look for in RUO and translational materials to ease the transition to the clinic including GMP and USP guidelines, what to consider when preparing for a clinical trial or a marketing application and common regulatory findings in cell therapy submissions that have the potential to delay approval. The panel will also discuss recent initiatives that have been put in place by regulators in different regions to accelerate cell therapy commercialization.
Jing He, PhD
Technical Application Scientist, Cell Biology, Life Sciences Solutions Group
Thermo Fisher Scientific
Optimization of your PSCs workflow
Background: Applications and general workflow of PSCs
- 高效和安全的重编程方法：当前各种重编程方法汇总，Cytotune 2.0介绍
Reprogramming with high efficiency and safety (Summary of current reprogramming methods, Introduction of Cytotune 2.0)
- PSCs培养的最佳体系：目前常见培养体系汇总， KSR-MS以及 Essential 8培养基介绍
Optimum PSCs culturing system: (Summary of current culturing system, Introduction of KSR-MS and Essential 8)
Characterization and differentiation of PSCs
Resources and tools
From Jul. 2012 to present, as a Technical Application Scientist of cell culture, protein expression and transfection related products in Life Sciences Solutions Group, Thermo Fisher Scientific.
Sep. 2007 - Jul. 2012, National Institute of Biological Sciences (NIBS, Beijing), Ph.D of Biochemistry and Molecular Biology. The main research area during the Ph.D period was stem cell, reprogramming and embryonic development.
Stephen Lin, PhD
Stephen Lin, PhD
California Institute for Regenerative Medicine
Initiatives to Advance Stem Cell Science and Medicine at California's $3 Billion Stem Cell Agency
The mission of the California Institute for Regenerative Medicine (CIRM) is to accelerate stem cell treatments to patients with unmet medical needs. With $3 billion in funding and 300 active programs, it is the world’s largest institution dedicated to helping people by developing cell therapies. To accomplish its mission, CIRM has funded a breadth of activities spanning from basic research to translation to clinical trials. In addition to individual grants, CIRM has created resources to help the stem cell community worldwide. Some resources promote research and drug development using stem cells. CIRM has established an iPSC Repository maintained by the Coriell Institute that is currently the largest publically accessible pluripotent stem cell bank in the world. CIRM also has a genomics research initiative that applies cutting edge sequencing and bioinformatics approaches to stem cell research and therapeutic development. Other resources established at CIRM promote the acceleration of stem cell discoveries into therapeutic development. CIRM is in the process of setting up a system of translational and clinical centers to help researchers move their discoveries into clinical development. This system includes 1) a Translating Center to support preclinical IND-enabling activities such as process development, safety testing, and cell manufacturing of stem cell therapy candidates, 2) an Accelerating Center that helps with clinical strategy, regulatory submission, and clinical trial management, and 3) an Alpha Clinics network that conducts clinical trials for stem cell related therapies. In total these resources are designed to catalyze the flow of stem cell discoveries to the clinic, which can come from both inside and outside of California. CIRM has supported 16 stem cell based clinical trials and the goal is to have 50. Beyond an overview of CIRM’s programs, this presentation will touch on perspectives around developing stem cell therapies and regulatory hurdles.
Stephen Lin is a senior science officer at California’s stem cell agency, CIRM. He joined the Agency in 2015 to oversee the its $32M initiative to create a repository of iPSCs from up to 3000 individuals covering both genetically complex and rare diseases, as well as a $40M genomics initiative that applies cutting edge genomics and bioinformatics approaches to stem cell research and therapeutic development. He is also the program lead on a newly created $15M preclinical research organization termed the Translating Center that focuses on preparing stem cell therapy candidates for clinical trials through support with process development, safety/toxicity studies, and manufacturing. From 2012 he had been a staff scientist and team lead at Thermo Fisher Scientific (formerly Life Technologies). Prior to that he was a scientist since 2006 at StemCells, Inc of California in the area of liver cell therapeutics. Dr. Lin received his Ph.D. from Washington University in St. Louis in 2002 under Jeffrey Gordon and did his postdoctoral research at Harvard University under Stanley Korsmeyer.
Rhonda Newman, PhD
Senior Staff Scientist
Thermo Fisher Scientific
Evolving your media system for modern stem cell applications
Culture systems for pluripotent stem cell (PSC) expansion enable generation of a nearly unlimited pool of cells for downstream differentiation, disease modeling, drug discovery, and therapeutic applications. While a number of PSC feeder-free medium systems exist, there are many challenges encountered by stem cell scientists across the PSC workflow in today’s applications. Here we sought to improve the robustness and versatility of traditional PSC culture medium systems identify and optimize critical medium components. Through assessment of over 65 different formulations, an optimum medium composition was identified which provides compatibility across the PSC workflow from somatic cell reprogramming, PSC expansion, downstream differentiation, as well as providing support in stressful applications such as gene editing. This system additionally provides versatility, allowing for every-other-day or weekend-free feed schedules and compatibility with a broad range of passaging reagents and matrices. Together this system provides a robust next-generation stem cell medium system for today’s challenging PSC workflow needs. For Research Use Only.
Dr. Newman joined Thermo Fisher Scientific in 2010 and is currently working on next generation stem cell culture and differentiation systems, enabling researchers to efficiently culture, expand, cryopreserve, and differentiate their stem cells to various cellular lineages. She received a Ph.D. at the University of Iowa from the lab of Dr. Madeline Shea. Subsequently, she completed postdoctoral training in the lab of Dr. Ken Prehoda at the University of Oregon, studying the role of intramolecular interactions in regulating cell signaling cascades in the process of asymmetric stem cell division.
Morrie Ruffin, PhD and Michael Werner, PhD
Advancing regenerative medicine and cellular therapies
The Alliance for Regenerative Medicine (ARM) is the preeminent global advocate for regenerative and advanced therapies. ARM fosters research, development, investment and commercialization of transformational treatments and cures for patients worldwide. By leveraging the expertise of its membership, ARM empowers multiple stakeholders to promote legislative, regulatory and public understanding of, and support for, this expanding field. ARM takes the lead on the sector’s most pressing and significant issues, including advocating for clear, predictable and harmonized regulatory and review pathways; enabling market access and value-based, favorable reimbursement policies; continued access to capital; addressing industrialization and manufacturing hurdles; developing and establishing industry-wide standards; and conducting key stakeholder outreach, communication and education.
Morrie Ruffin has more than 20 years of experience in the Biotech and Healthcare industries. He is the co-founder and managing director of the Alliance for Regenerative Medicine (ARM). He is also the managing partner of Adjuvant Partners, a life sciences advisory practice working with product developers and major medical centers advancing programs in cell and gene therapy. Prior to joining Adjuvant in 2008, he was the Chief Executive Officer of LifeTech Innovations. From 1994-2006, Mr. Ruffin was Executive Vice President of Capital Formation and Business Development at the Biotechnology Industry Organization (BIO), the largest trade organization representing the biotech and drug development industries. Joining BIO in 1994 as one of its original employees, Mr. Ruffin was responsible for building the organization's global business development and investor outreach programs focused on helping companies raise capital and identify strategic partnering and licensing opportunities. This BIO business development franchise is now the largest in the world, with operations in the US, Europe, and Japan.
In addition to his business development work at BIO, Mr. Ruffin was responsible for leading the industry's capital formation advocacy efforts with a focus on economic incentives to promote investment in early stage biotech and med-tech businesses. He is also a founder and board member of the Interoperable Informatics Infrastructure Consortium (I3C), an international standards setting body for the bioinformatics industry.
Prior to joining BIO, Mr. Ruffin worked for US Senator Arlen Specter for five years as his senior legislative assistant. Prior to that, he spent approximately five years working in varying capacities, including policy analyst at Systems Planning Corporation International and the Center for Strategic and International Studies. Mr. Ruffin received his MA in International Studies & Economics from the Johns Hopkins School for Advanced International Studies (SAIS) and his BA from the University of Virginia.
Michael Werner is a partner in Holland & Knight's Washington, D.C. office and co-chair of the firm's Healthcare & Life Sciences Industry Team. He has almost three decades of healthcare law, regulatory, reimbursement, and lobbying experience in Washington. He focuses on issues affecting biotechnology and pharmaceutical companies, medical research and research institutions, physicians and patients. His specific areas of knowledge include FDA regulations regarding drug/biological product review, approval, and distribution; regulation of cell therapy, gene therapy, tissue engineering and regenerative medicine products; IRB review, informed consent, and other clinical trial issues; regulation and reimbursement of biosimilars; Medicare reimbursement strategy and issues; as well as conflicts of interest and other bioethics issues arising from research and uses of new technologies.
Mr. Werner is the co-founder and Executive Director of the Alliance for Regenerative Medicine, a Washington, DC-based organization comprised of over 240 member companies, clinical centers, patient advocacy groups and other organizations world-wide whose mission is to advocate for regulatory and reimbursement policies that will advance regenerative medicine research and product development.
Before joining Holland & Knight, he was president of The Werner Group, a Washington, D.C.-based firm that provided lobbying, regulatory, and bioethics consulting services for biotechnology and pharmaceutical companies, physicians, health plans, investors, and patient advocacy groups. Prior to founding The Werner Group, he was chief of policy for the Biotechnology Industry Organization (BIO), representing over 1000 biotechnology companies in the U.S. and other countries. In that role, he was responsible for virtually all major issues affecting biotech companies including: drug/biologic evaluation and review by FDA; CMS policies and reimbursement, Medicare, intellectual property, stem cell research and bioethics issues.
Mr. Werner was senior healthcare advisor to U.S. Senate Majority Leader George Mitchell, a congressional investigator for the U.S. Senate Special Committee on Aging and senior advisor to Maryland Governor William Donald Schaefer.
Mr. Werner is a frequent media commentator and has appeared in The Wall Street Journal, Science, Scientific American, and The Washington Post. In 2013 and 2014, he was named one of the Top 50 Global Stem Cell Influencers by Total BioPharma. He was also recently named to the US National Academies of Science, Engineering and Medicine’s Forum on Regenerative Medicine.
He is a heavily sought-after speaker for meetings and conferences, and the author of over 50 published articles. Michael is co-author of the Bloomberg BNA portfolio series “Life Sciences Compliance: a Pre-market and Post-market Portfolio”. His article “Don’t Edit the Human Germ Line” appeared in the March 2015 issue of Nature.
Lay Teng Ang, PhD
Senior Research Fellow
Genome Institute of Singapore
Precise generation of desired human cell-types from embryonic stem cells
My research program focuses on building the technology to generate diverse human cell-types for application in regenerative medicine and drug toxicology testing. While embryonic stem cells (ESCs) have the potential to generate thousands of distinct human cell-types, this vast array of lineage choices has made it difficult to efficiently differentiate ESCs towards any single desired fate. To more accurately guide ESC differentiation, we have delineated comprehensive roadmaps that describe how ESCs are diversified into a wide variety of endodermal and mesodermal cell-types. These roadmaps have enabled the successful generation of human liver cells and bone progenitors that can engraft in mouse models and respectively regenerate human liver tissue or bones in vivo; as such these ESC-derived tissue progenitors serve as potential sources of human cells for regenerative medicine or drug toxicology testing.
Lay Teng is currently a Visiting Assistant Professor at Stanford University and a Senior Research Fellow at the Genome Institute of Singapore, A*STAR. She received her B.A. (Honours) in Bioengineering from the National University of Singapore and her Ph.D. from the University of Cambridge under an A*STAR Scholarship. Lay Teng has eleven years of research experience and dedicated the past nine years working with human embryonic stem cells (hESCs) and their differentiation towards endodermal and mesodermal cells. Her research has been supported by two ETPL Gap Funding Grants from A*STAR (as PI and co-PI, respectively) and has led to manuscripts in Cell and Cell Stem Cell (as corresponding author). She further led the commercialization effort to position her technology as a research kit, which is now sold worldwide by Thermo Fisher Scientific, Inc. Her ultimate goal is to develop new human cell types for use in regenerative medicine and drug toxicology testing.
Randolph Ashton, PhD
University of Wisconsin Madison
Derivation of a Spectrum of Regional Motor Neuron Phenotypes for Hindbrain and Spinal Cord Regenerative Medicine
The central nervous system (CNS) is a conglomerate of diverse, interconnected tissues that each contain cell phenotypes specific to their distinct anatomical region. Recent studies have demonstrated that CNS cells derived from human pluripotent stem cells (hPSCs) must be of the appropriate regional phenotype to model tissue-specific disease pathologies in vitro as well as produce a regenerative effect upon transplantation. Yet, only a limited number of regional CNS phenotypes can be derived from hPSCs due to the complex regimen of developmental cues necessary to effectively instruct region-specific neural differentiation.
Here, we present a chemically defined protocol for deriving forebrain neural stem cells (NSCs) from hPSCs with greater than 90% efficiency in under 6 days (Lippmann et al. Stem Cells 2014). Also, we have successfully deciphered the regimen of developmental cues that govern differentiation of hPSCs into NSC phenotypes specific to any diverse hindbrain or spinal cord region (Lippmann et al. Stem Cell Reports 2015). These NSC cultures can be differentiated into a spectrum of respective regional hindbrain and spinal cord motor neuron phenotypes, which can be matured to fire action potentials and innervate skeletal muscle fibers in vitro.
Hence, our chemically defined protocols vastly expand the diversity of regional CNS phenotypes that can be derived from hPSCs. They enable access to the hundreds of different regional motor neuron phenotypes present in the human hindbrain and spinal cord, which are the sole means of transmitting efferent signals from the CNS to peripheral tissues including skeletal muscles that provide motor function. Our findings have significant implications for modeling degenerative disorders that target hindbrain and spinal cord motor neurons (e.g. Amyotrophic Lateral Sclerosis) and developing regenerative cell therapies for paralysis.
Randolph S. Ashton received his B.S. from Hampton University (Hampton, Virginia, 2002) and Ph.D. from Rensselaer Polytechnic Institute (Troy, NY, 2007) in Chemical Engineering. During graduate studies under Prof. Ravi Kane, he researched how engineering biomaterials at the nanoscale could regulate the fate of adult neural stem cells. He continued to pursue his interest in stems cells and tissue engineering as a California Institute for Regenerative Medicine and a NIH postdoctoral fellow at the University of California Berkeley’s Stem Cell Center in the lab of Prof. David Schaffer. In 2011, he was appointed to a faculty position in the Wisconsin Institute for Discovery at the University of Wisconsin–Madison as an Assistant Professor of Biomedical Engineering. The goal of Dr. Ashton’s research is to provide novel regenerative therapies to treat CNS diseases and injury. His lab is currently developing scalable protocols to generate region-specific central nervous system tissues from human pluripotent stem cells (hPSCs). They also meld state of the art biomaterial approaches with hPSC-derived neural stem cells to engineer brain and spinal cord tissue models in vitro. Among his awards and honors, Dr. Ashton was named the 2016 Young Faculty Investigator Awardee by the Regenerative Medicine Workshop at Hilton Head, a 2015 Emerging Investigator by Chemical Communications, and a 2013 Rising Star by the Biomedical Engineering Society’s Cellular and Molecular Bioengineering Special Interest Group. Also, he has been awarded a Burroughs Wellcome Fund Innovation in Regulatory Science Award, a Draper Technology Innovation Award from the Wisconsin Alumni Research Foundation, a Basic Research Award from the UW Institute for Clinical & Translational Research. His research is also supported by grants from the NIH and EPA.
Pau Sancho Bru, PhD
Institut D'Investigacions Biomediques August Pi I Sunyer (IDIBAPS
Directed differentiation of induced pluripotent stem cells to hepatic stellate cells
In healthy liver, quiescent hepatic stellate cells (HSCs) participate in the homeostasis of extracellular matrix and store vitamin A. After injury, HSCs activate and participate in the wound-healing response, producing extracellular matrix components and eventually fibrosis. We have developed a protocol to direct the differentiation of human induced pluripotent stem cells (iPSC) to HSCs. The final HSC–like population was enriched in PDGFRß positive cells and expressed key HSC markers at similar levels than primary HSC. Whole genome transcriptomic analysis revealed that PSC-derived population displayed an intermediate phenotype between activated and quiescent HSCs. Functional analysis showed that PSC-derived HSC-like cells responded to injury mediators and accumulated retinyl esters into lipid droplets. These findings show that we have generated functional HSC-like cells from iPSC, which may have potential for in vitro and biomedical applications.
Pau Sancho-Bru received his Ph.D in 2006 from the University of Barcelona. In 2007 he joined the Stem Cell Institute Leuven at the KULeuven, Belgium as a postdoctoral researcher. In 2012 he was appointed Researcher in IDIBAPS. Pau Sancho-Bru is Principal Investigator at the Liver Injury and Repair Group and Researcher at IDIBAPS. His group is focused on understanding the mechanisms governing liver injury and regeneration and particularly the role of hepatic stellate cells and liver stem/progenitor cells in wound-healing response. One of the main research interests of his group is developing in vitro systems for disease modeling and assessing the potential of stem cells for biomedical and biotechnological applications.
Vijay Chandrasekar, PhD
Elucidating the physiological function of cellular PrPC using human iPS cel
PrPc is a conserved lipid-raft associated, GPI-anchored cell membrane glycoprotein. Misfolding of cellular PrPc into the pathogenic PrPSc results in Prion disease, an untreatable and fatal neurodegenerative disorder. Prion induced neurotoxicity is preceded by impairment in metabolism of cholesterol and other lipids which are major component of lipid-rafts in affected neurons. Lipid-rafts deregulation has been implicated in diseases like Prion and AD, the mechanism remains unclear. Understanding the function of cellular PrPc may shed light on such pathological mechanisms. Towards this goal, we utilize a human induced pluripotent stem (iPS) cell model system. We generated isogenic PrP knockout (KO) human primary fibroblasts in order to reprogram them into PrP-KO-iPS cells and human neurons. Metabolomics and RNAseq analysis of these PrP-KO human cells show dysregulation in key CNS pathways like glycerophospholipid and cholesterol metabolism. Here we present a systems biological approach combining RNAseq and metabolomics to understand the functional molecular network of PrPc. The knowledge of key pathways in which PrPc has important implications could aid in the targeted therapy for prion disorders
Vijay Chandrasekar was born in the southern part of India called Tamil Nadu. He did his Master studies in Microbial Gene technology in Madurai Kamaraj University. Following that he worked as a Research associate in the premier institute in India called India Institute of Science, where he worked on Protein biochemistry and Crystallography, where he was successfully involved in structure determination of several viral proteins resulting in good publications. For his PhD, Vijay moved to Switzerland in 2010, where he did his doctorate studies in Molecular Neuroscience on “Characterization of microRNA and transcription factor gene network in cocaine induced neuroplasticity” in the University of Fribourg, Switzerland. His studies established the role of miRNAs in the addiction and neuronal plasticity induced by drugs of abuse in the brain for the very first time and resulted in several highly cited publications. After his PhD, Vijay moved to US to pursue his first Postdoctoral fellowship in Columbia University Medical Center (CUMC), NYC, under the renowned scientist Prof. Tom Maniatis. During which time, he had his own Helmsley Stem cell starter grant in CUMC for his studies on ALS disease using human iPS cells. Vijay worked on two interesting projects (a) HILO-RMCE based generation of iPS cells for studying C9orf72 mutation in ALS. (b) "Identifying the role of miRNAs in astrocyte dependent, non-cell autonomous motor neuron toxicity". He successfully initiated footprint free reprogramming for in vitro disease modelling using iPS cells. Vijay is currently pursuing his second postdoctoral Scientist position in the Institute of Neuropathology, University Hospital Zurich, Switzerland, working under Prof. Adriano Aguzzi on “Establishing stem cell based systems ES and iPS cells for studying Prion mutation in CJD”. For the past almost 6 years, he has been working on mouse and the human stem cells both ES and iPS, to derive neurons and other cell lineages for disease modelling, characterization and drug screening assays using variety of state-of-the-art methods like survival, morphometric measurements, RNAseq and proteome analysis. Vijay has established the model systems in CUMC and in the University Hospital, Zurich as well as in our collaborative labs in Zurich.
Davide Danovi, PhD
Director, HipSci Cell Phenotyping
Centre for Stem Cells and Regenerative Medicine, King's College London
‘HipSci' and the ‘Stem cell hotel’: innovative platforms for collaborative phenotyping
We work in the framework of the Human Induced Pluripotent Stem Cells Initiative (HipSci) project, funded by the Wellcome Trust and MRC. We provide a dedicated laboratory space for collaborative cell phenotyping to study how intrinsic and extrinsic signals impact on human cells to develop assays for disease modeling and drug discovery and to identify new disease mechanisms.
Davide Danovi holds an MD from University of Milan and a PhD in Molecular Oncology from the European Institute of Oncology where he demonstrated the causative role of the HdmX protein in human cancer. He completed his postdoctoral training working with Prof. Austin Smith and Dr. Steve Pollard at the University of Cambridge and at University College London where he developed a screening platform to isolate compounds active on human neural stem cells from normal or brain tumour samples. Prior to his current role, he worked as principal scientist at a novel biotechnology company founded to isolate drugs for regenerative medicine using innovative stem cell technologies.
Chris Denning, PhD
Professor in Stem Cell Biology
University of Nottingham
Disease modelling in pluripotent stem cell-derived cardiomyocytes
Over the last 15 years, human pluripotent stem cell (hPSC) technologies have progressed from academic curiosities into tools with the promise to underpin commerce, leading to real progress in understanding of disease, improving drug safety and providing novel routes to clinical translation. With an emphasis on the heart, this presentation will discuss our progress in producing models of genetic disease by reprogramming somatic cells into human induced pluripotent stem cell (hiPSC). This includes various conditions such as long QT syndrome, Duchenne muscular dystrophy and CPVT, which affect the function and / or structure of cardiomyocytes. We will show how the Cas9/CRISPR system is being used to produce defined sets of polymorphisms in the ADRB2R and GRK5 loci, which encode proteins that underpin β2-adrenoceptor signaling. These polymorphisms reflect the genotypes in the patient population and we will present early data on how these changes may influence receptor density, internalization and both receptor and heart function. Since these panels of hiPSC and engineered lines can now be created with relative ease, bottlenecks of scaled culture, differentiation and phenotyping are becoming a considerable issue. Thus, we have developed an automation suite that includes a bespoke robotic platform to culture and differentiate hPSCs at scale into cardiomyocytes. Into this suite, we have incorporated high content platforms that allow assessment of structure (confocal plate reader imaging) and function (mitochondrial activity, contractility and electrophysiology). Despite these advances, numerous challenges remain, such as incomplete epigenetic reprogramming of hiPSC relative to hESCs and insufficient levels of expression of key ion channels, which need to be considered for the applicability of these models in biomedical application.
Chris Denning is a Professor in Stem Cell Biology, with particular interests in cardiomyocyte (heart cell) differentiation of human embryonic and induced stem cells for use in drug screening and in production of new in vitro models of genetic-based cardiovascular disease. This includes manipulation of the genome using transgenic and nuclease-mediated gene targeting technologies (including Cas9/CRISPR). In parallel, Chris has also focused on optimisation of the culture environment and robotic culture to allow fully automated scale-up and high throughput screening, using high content electrophysiology and imaging.
Faranak Fattahi, PhD
Memorial Sloan-Kettering Cancer Center and Weill Cornell Medicine
Human pluripotent stem cells for the study of enteric neuropathies
The human enteric nervous system (ENS) is derived from the neural crest and represents a complex network of ~500 million neurons with dozens of distinct neurotransmitter and hormone subtypes essential for gastro-intestinal (GI) activities. Despite the significance of the human ENS and its involvement in a wide range of GI disorder, there is very little known about its biology due to the lack of accessible tissue. Directed differentiation of human pluripotent stem cells (hPSCs) offers and alternative approach for the derivation of distinct, functional cell types that can be utilized in a broad range of basic and translational applications. We have recently succeeded in derivation of the ENS lineages from hPSCs and applied them for cell therapy and drug discovery in Hirschsprung disease, which is the most common developmental disorder of the ENS. These studies set the stage for future investigations of the human ENS biology and a better understanding of disease mechanisms and advancement of therapeutic interventions for enteric neuropathies.
Faranak Fattahi has been a graduate student in the laboratory of Dr. Lorenz Studer at Memorial Sloan Kettering Cancer Center. During her PhD, she focused on development of new strategies to derive lineages of the peripheral nervous system (PNS) from human pluripotent stem cells (hPSCs) and demonstrated their potential for disease modeling, drug discovery and regenerative medicine. Upon completion of her graduate training, she will join University of California San Francisco as a Sandler Faculty Fellow to launch her research program on the application of hPSCs for the study of the human PNS in health and disease.
Ben Fryer, PhD
Team Leader, Processing/Manufacturing, Heart Regeneration Program
University of Washington School of Medicine
Transitioning to the clinic from proof of concept research: Challenges associated with converting research materials and methods to a clinical grade product
Years of significant effort can go into generating key proof of concept data to support moving bench research to clinical development. Yet, much work remains in order to transition from POC to clinical trials for a cell therapy. Raw materials, processes, and cell banks may all require changes to meet regulatory guidances and more stringent specifications for treating a patient. We will discuss the practical aspects of developing a cell therapy to treat heart disease.
Ben Fryer is the team leader for CMC-Cell Manufacturing and Processing at the University of Washington School of Medicine’s Heart Regeneration Program (HRP). Ben joined the HRP in January 2015. The program was started by Chuck Murry and is currently in pre-clinical development of a therapy to treat heart disease using cardiomyocytes generated from hES cells.
Prior to joining HRP, Ben worked at Janssen R&D’s internal diabetes venture (BetaLogics) from 2006 to 2014. BetaLogics’ mission was to treat insulin dependent diabetes with a combination islet-like tissue product generated from hES Cells and delivered via an immune-isolation device. Ben managed an internal research team and several external partnerships dedicated to finding a defined and scalable method to manufacture pancreatic beta islets for the treatment and cure of diabetes.
Ben earned a BA from Colorado College and his PhD in pharmacology from the University of Pennsylvania. As a post-doc he studied Hypoxia Inducible Factor in cancer and stem cell development in the Howard Hughes Laboratory of M. Celeste Simon at the University of Pennsylvania. Prior to graduate school he worked for several small biotech / pharmaceutical firms in the Denver-Boulder area supporting discovery research and pre-clinical & clinical development of small and large molecule therapies for cancer.
Ben is an inventor on several pending patents for Janssen/Johnson&Johnson, including stirred tank bioreactor based suspension expansion and differentiation processes and a product currently marketed by Thermo-FisherTM as Nunclon-Vita.
William Hendricks, PhD
Instructor in Neurology
Massachusetts General Hospital | Harvard Medical School
Human PSC-based disease modeling to study X-linked Dystonia-Parkinsonism
The isolation of human embryonic stem cells (hESCs) and the discovery of human induced pluripotent stem cell (hiPSC) reprogramming have sparked a renaissance in stem cell biology, in vitro disease modeling, and drug discovery. In general, hPSC-based disease models are well-suited to study genetic variation. Studies commonly compare patient-derived hiPSCs, e.g., with a disease-causing genetic mutation, and (age-matched) control subject-derived hiPSCs, typically differentiated to the disease-affected cell type, e.g., neurons. A major caveat of this disease-modeling strategy is the variability of differentiation propensities and phenotypic characteristics, even in hPSCs derived from the same donor. Still, even if the cellular phenotype of a given mutation is strong and highly penetrant, it may be lost due to confounding effects of differences in genetic background of unrelated hPSC lines. A very powerful approach to overcome this hurdle is to use custom-engineered endonucleases, such as CRISPR/Cas9 that enable precise and programmable modification of endogenous hPSC genomic sequences. In our lab we use hPSC-based disease modeling to study the neurological movement disorder dystonia, in particular X-linked Dystonia Parkinsonism (XDP). In this talk I will show how we use hPSC-based disease modeling in combination with CRISPR/Cas9 gene editing, to elucidate the underlying molecular pathogenesis of XDP. I will also discuss some of the potential problems one might face using hPSC-based disease modeling in combination with gene editing.
After receiving his PhD in Neuroscience at the VU University Amsterdam in the Netherlands in 2008, Dr. Hendriks joined the lab of Dr. Paola Arlotta at the Center for Regenerative Medicine of Massachusetts General Hospital (MGH) in Boston to study neuronal development focusing on neuronal differentiation of human pluripotent stem cells. In 2011, Dr. Hendriks joined the Harvard Stem Cell Institute (HSCI) iPS Core facility with Dr. Chad Cowan at Harvard University in Cambridge, where initially he worked on developing and implementing foot-print free somatic cell iPSC reprogramming methods. Dr. Hendriks also initiated and managed the hPSC genome editing service for 2 years at HSCI before moving to his current position as a Harvard Medical School Instructor in Neurology at the MGH Collaborative Center for X-Linked Dystonia Parkinsonism.
Lachlan Jolly, PhD
ARC DECRA Fellow
The University of Adelaide
Use of Stem Cell models to assess Genetic Change Underlying Neurodevelopmental Disorders
The study of human genetic neurodevelopmental disorders (NDDs) is complicated by the inaccessibility of the relevant tissue for study: it is extremely rare to obtain post-surgical brain samples from patients, and the origins of disease often occur in-utero. The technologies of ex-vivo stem cell culture is a valuable tool to study embryonic development. Derived from the embryo and cultured under appropriate conditions, both embryonic stem and committed neural stem cells display multipotent features of the tissue they are derived from, and are licensed in-vitro with the properties of unlimited self-renewal. The subsequent differentiation of these cells recapitulate many aspects of in-vivo development and can be used to model embryonic brain development. Because of these attributes, and the ease of genetic and other manipulations afforded in-vitro, we have employed these models to assess the effects of gene mutations that underlie neurodevelopmental disorders. We have been able to assign neurodevelopmental functions to newly discovered novel genes, test the pathogenicity of variants of unknown significance, and identify aspects of brain development likely affected in patients harbouring deleterious mutations.
Dr Lachlan Jolly completed his PhD at the University of Adelaide, South Australia, in 2010 during which he applied neural differentiation of embryonic stem cells to study the earliest stages of brain development. He next joined the Neurogenetics Research Program headed by Professor Jozef Gecz for post-doctoral training at the Women’s and Children’s Hospital in Adelaide (SA Pathology). His application of various neural cell culture models of brain development resulted in the discovery of several new genetic causes of neurodevelopmental disorders. Dr Jolly now leads his own research group at the University of Adelaide, focusing on the roles of the Nonsense Mediated mRNA Decay (NMD) pathway, and the genes USP9X, HCFC1, and PCDH19 in normal and pathological processes of brain development and function. He is currently an Australian Research Council DECRA Fellow.
Leo Kurian, PhD
Junior Research Group Leader
Building embryonic lineages
The human body is composed of about 200 different cell types. The identity and function of these distinct cell types are precisely programmed by the regulatory networks encoded in the 3 billion base pairs of DNA that constitute the human genome. While 60% of our genome is transcribed, less than 2% of it is translated to proteins. In contrast to previous assumptions, this suggests that a significant majority of the regulatory information from the genome functions as RNAs, termed non-coding RNAs. Emerging evidences suggest that a substantial portion of these non-coding transcripts control myriad biological processes ranging from development to disease, establishing the vital role played by these RNA regulatory elements. In addition, these molecules are regulated by RNA binding proteins at the functional level. We investigate how RNA regulatory elements program cellular identities during cardiac development, aging, and regeneration.
Leo Kurian completed his basic education in chemistry followed by a Master’s degree in biotechnology in India. He obtained his PhD in genetics from the University of Cologne. He spent his post-doctoral years in the Belmonte lab at the Salk Institute and in the Yeo lab at UCSD (both in San Diego, California), where he established stem cell-based models to study programming and reprogramming of cell-fate decisions. In 2014, he established an independent group, supported by the NRW Stem Cell Network, to study the regulatory basis of cardiac development, aging and regeneration at the University of Cologne.
Willie Lin, PhD
Chief Executive Officer
Meridigen Biotech Co., Ltd.
Human umbilical cord mesenchymal cells and the treatment of bronchopulmonary dysplasia
The human umbilical cord is a promising abundant source of mesenchymal stem cells (MSCs). Compare to other MSCs, the advantages of human umbilical cord MSCs (hUC-MSCs) are easily accessible, short amplification time, high proliferation rate, lower immunogenicity, and abundance. Many studies have shown that hUC-MSCs have the ability to differentiate into three germ layers, to accumulate in damaged tissue or inflamed regions, to promote tissue repair, and to modulate immune response. Thus, they are attractive human cell–based therapies for the treatment of bronchopulmonary dysplasia, chronic obstructive pulmonary disease, Ischemic stroke, neuron degenerative diseases, autoimmune diseases, etc.
Dr. Willie Lin received his bachelors of science in Animal Sciences from Tunghai University in Taiwan followed by a doctorate in Biochemistry from Kansas State University in the USA. He is currently serving as the Chief Executive Officer of Meridigen Biotech, Co., Ltd based in Taiwan. Dr. Lin’s professional experience has included: Adjunct Associate Professor of the Master Program in Technology management at Fu Jen Catholic University, Chairman and President of Microbio Co., Ltd and Fountain Biopharma, Inc, President and Chief Operating Officer at UniMed Venture management, Inc, Associate Vice President of the Strategy and Planning Department at China Chemical and Pharmaceutical Co., and Associated Vice President in the Overseas Business Department for the China Development Industrial Bank.
Jeanne Loring, PhD
The Scripps Research Institute, Center for Regenerative Medicine
Stem Cells for Regeneration and Rescue.
Most discussions about pluripotent stem cells center around their promise for regenerative medicine. The most remarkable quality of these cells is their ability to turn into all of the cell types in the body. This amazing power is driving all of the projects in our lab that range from treatment of human disease to rescue of endangered species. One of our projects is development of an autologous cell replacement therapy for Parkinson’s disease (PD). We have used non-integrating Sendai virus to reprogram skin cells into induced pluripotent stem cells (iPSCs) from ten people afflicted with PD. Since death of dopamine neurons in the brain leads to the motor symptoms of PD, we have developed methods for generating dopamine neurons from iPSCs and plan to use these cells for autologous transplants, which should eliminate the need for immunosuppression. On the other end of the spectrum, we are using iPSCs in an effort to save endangered species. We are generating iPSCs from fibroblasts stored in the Frozen Zoo® at the San Diego Wild Animal Park. We are focusing on the Northern White Rhino, which will be extinct in our lifetimes. Only 3 members of this species remain alive. But there are fibroblasts from 12 individuals stored in the Frozen Zoo, and we have made iPSCs from two so far. We are generating iPSCs from the rest, and plan to differentiate them into gametes- this has been achieved for mice, but we don’t expect it to be easy for rhinos. If we succeed, and can make fertilized embryos in the lab, there are 6 females from the closely related Southern White Rhino species already at the zoo who will serve as surrogate mothers. The astounding power of pluripotency has opened doors to new ideas; we’re just getting started.
Jeanne F. Loring is a Professor and the founding Director of the Center for Regenerative Medicine at The Scripps Research Institute in La Jolla. Her research team focuses on large-scale genomic and epigenetic analysis of human pluripotent stem cells (hPSCs), with the goal of ensuring their effectiveness and safety for cell therapy. Her lab is developing stem cell-based therapies for Parkinson’s disease and multiple sclerosis, and investigates the underlying causes of autism using patient-specific stem cells. With the San Diego Zoo, her lab is developing a "zoo" of induced pluripotent stem cells from endangered species to aid in their conservation.Dr. Loring serves on many scientific and bioethics advisory boards, including the Merck KGaA Bioethics Advisory Panel (Germany) and the scientific advisory boards for Genea Biocells, Inc. (Australia), Kadimastem, Inc.(Israel), Coriell's NIGMS Human Genetic Cell Repository, the National Center for Biomedical Glycomics, the NIMH Repository & Genomics Resource (Rutgers), and the Heart Regeneration Program (U. Wash.). She was a member of the Panel on Global Assessment of Stem Cell Engineering (NSF, NIST, and NIH) and the Panel on Review of the Material Measurement Laboratory at NIST (The National Academies). She is frequently quoted in major newspapers, appears on television and in documentary features, and gives many public lectures about science and society. She is particularly concerned with the dangers of unregulated stem cell treatments in the US and other countries.
Anish Sen Majumdar, PhD
CSO & Executive VP
First Allogeneic Mesenchymal Stromal Cell Product Approved in India for Buerger’s Disease – An Unmet Medical Need
Buerger’s disease, commonly known as Thromboangiitis obliterans, is a non-atherosclerotic, segmental inflammatory disease that can affect the small and medium-sized arteries of young people with a history of heavy tobacco use. Its prevalence among all patients with peripheral arterial disease varies widely in different regions of the world. Critical limb ischemia (CLI) is a severe form of the disease that results in acute rest pain and non-healing ischemic skin ulcers and gangrene of the lower extremity which can results in limb amputation if left untreated. Bone marrow derived MSC (BMMSC) are known to possess strong immunomodulatotory properties, promote angiogenesis and tissue regeneration through paracrine activity. Using a patented pooling technology of BMMSC from different healthy donors, we have developed an allogeneic BMMSC product, Stempeucel. Stempeucel is manufactured in a GMP facility and cryopreserved as an off-the-shelf product . A series of preclinical studies were conducted to establish the safety  and efficacy profiles of the pooled BMMSC population. More importantly, results obtained from our phase I/II clinical trial in CLI patients demonstrated the safety of Stempeucel in humans . A large phase II was recently completed with two different doses of Stempeucel. The cells were injected intramuscularly at multiple locations around the calf muscle and also around the ulcer. Analysis of the data six months after Stempeucel administration revealed statistically significant improvements in patients for both the primary clinical end points i.e., relief of rest pain and ulcer healing in the target limb as compared to the control group of patients . In addition, ankle brachial pressure index (ABPI) also showed significant improvement in the same cell dose group, suggesting improved blood flow in the limb. As such, Stempeucel has been approved by the regulatory authorities in India for its use in a limited number of “No Option” Buerger’s disease patients.
Dr. Anish S Majumdar is the Chief Scientific Officer & Executive VP of Stempeutics Research in Bangalore, India, since 2010. He has been working in the field of stem-cell based research and product development for more than 20 years. Dr. Majumdar received his Ph.D. in Biochemistry/Immunology from the University of Calcutta, India. He pursued postdoctoral research at the University of Pennsylvania and subsequently joined Stanford University as a Research Scientist.
He has worked at Indian and US based biopharmaceutical/cell therapy companies in various positions: Among the US companies are BD Biosciences, Aventis, and Geron Corporation. Dr. Majumdar joined Geron Corporation in 1998 and became Senior Director, Cell Therapy Research in 2005. In 2007, he relocated to India where he joined Reliance Life Sciences, Mumbai as Vice President of Stem Cell Research & Regenerative Medicine. He has published nearly 80 research articles, reviews, and book chapters in acclaimed journals and is a co-inventor of 70+ international patents. Dr. Majumdar’s research expertise spans from the biology, immunology and therapeutic applications of mesenchymal stem cells (MSC), differentiation of human embryonic stem cells (ESC), dendritic cells, and T-cells for developing anti-cancer vaccines. Currently, the use of MSCs for various therapeutic applications in regenerative medicine is his focus at Stempeutics, and has met with strong success.
He is a member of the Industry Committee of the ISSCR, Federation of Indian Chamber of Commerce, and Industry (FICCI) for the Biotechnology Chapter as a stem cell expert. In 2013, Dr. Majumdar was elected as Vice President for Asia region of the ISCT. In this position, he represents India and Asia, and he is committed to promoting the use of stem cell based therapeutics for diseases with unmet or urgent medical need.
Nuria Montserrat, PhD
Junior Group Leader
Institute of Biotechnology and Engineering
Tissue engineering with human pluripotent stem cells
One of the ultimate goals in Regenerative Medicine is the generation of pluripotent stem cells (PSCs) directly from somatic cells obtained from patients. Although major findings in the definition of in vitro differentiation protocols allowing for the derivation of several somatic cell lineages has been reported progresses on obtaining certain defined populations has faced major obstacles. One of the major problems associated with the in vitro differentiation arises in tissues presenting complex tridimensional (3D) structures, such as the case of cardiac, hepatic or renal lineages. Lately, it has been described that complex differentiation strategies might benefit from the establishment of 3D culture systems. The possibility to develop 3D dimensional self-organized tissues, so called organoids, has opened new venues in the generation of protocols dictating human PSCs differentiation. As tissue-derived organoids are compromised by multiple cell types recapitulating part of organ structure and function the possibilities to understand human development are facilitating the development of massive platforms for drug screening and personalized medicine. In this talk, we will summarize part of our recent work in heart and kidney differentiation using hPSCs highlighting potential applications when combined with organ decellularization and 3D bioprinting
Dr. Montserrat has undergraduate training in Spain, Switzerland and France, and postgraduate training in Spain and the US. Her research career started early in 2008 at the Center of Regenerative Medicine of Barcelona (CMRB), under the direction of Dr. Izpisua Belmonte. There, she led and participated different projects involving the generation and banking of iPS lines (Cell Stem Cell, Nature Protocols, Nature Methods, Protein and Cell, Nature, among others), and set up safe strategies using specific transcription factors determinant for lineage specification (GATA3) for somatic reprogramming (Cell Stem Cell, 2013). She also collaborated in other projects aimed to characterize the genomic integrity of human iPSCs as well as in the differentiation of iPSCs towards germ cells, neural cells, endothelial cells, retinal cells or blood cells. These studies were published in Stem Cells 2011; Nature 2012; Nature Methods 2012, Nature Communications 2013, Protein and Cell 2013, Nature Communications 2014. In the same manner, Dr. Montserrat participated in the generation of platforms for the study of disease progression and compound screening for therapy by means of human iPSCs [Nature 2012, Nature Communication 2014]. Moreover, her interest on organ regeneration provide new knowledge for the generation, for the first time, of kidney organoids, suitable for the study of hiPSCs differentiation towards renal lineages and compound screening for therapeutic purposes [Nature Cell Biology (2013)]. Recently she has identified, for the first time, how the reactivation of endogenous regenerative programs that are dormant in adult murine heart can be reactivated and elicit heart regeneration [Cell Stem Cell, 2014]. Dr. Montserrat research also benefits from the use of bioengineering strategies (organ decellularization and 3 D bioprinting) for human pluripotent stem cells differentiation, with specific focus in heart (Biomaterials, 2016) and kidney. The European Research Council (ERC-ERC Starting Grant), and Spanish national programs and networks support Montserrat research. Currently, Dr. Montserrat is group leader at the Institute of Bioengineering of Catalonia (IBEC), in Barcelona. Her research is focused in the study of molecular programs sustaining reprogramming and differentiation, with special focus in mesodermal derived tissues as kidney and heart.
Cory Nicholas, PhD
Co-Founder and Chief Scientific Officer, Neurona Therapeutics
Assistant Professor, Adjunct, University of California, San Francisco
Transplanted human stem cell-derived interneuron precursors mitigate mouse bladder dysfunction and central neuropathic pain after spinal cord injury
Neuropathic pain and bladder dysfunction represent significant quality of life issues for many spinal cord injury patients. Loss of GABAergic tone in the injured spinal cord may contribute to the emergence of these symptoms. Previous studies have shown that transplantation of rodent inhibitory interneuron precursors from the medial ganglionic eminence (MGE) enhance GABAergic signaling in the brain and spinal cord. Here we look at whether transplanted MGE-like cells derived from human embryonic stem cells (hESC-MGEs) can mitigate the pathological effects of spinal cord injury. We find that six months after transplantation into injured mouse spinal cords, hESC-MGEs differentiate into GABAergic neuron subtypes and receive synaptic inputs, suggesting functional integration into host spinal cord. Moreover, the transplanted animals showed improved bladder function and mitigation of pain-related symptoms. Our results therefore suggest that this approach may be a valuable strategy for ameliorating the adverse effects of spinal cord injury.
Find the published paper online at: target="_blank">http://www.cell.com/cell-stem-cell/abstract/S1934-5909(16)30267-3
Dr. Cory Nicholas is Chief Scientific Officer at Neurona Therapeutics. Prior to launching Neurona, Dr. Nicholas was a faculty member in the Department of Neurology at the University of California, San Francisco where his research program was focused on elucidating the ontogeny of human cortical interneurons. Using embryonic brain development as a blueprint, Dr. Nicholas pioneered methods to derive interneuron precursors from human pluripotent stem cells and developed transplantation cell-based therapies for multiple animal models of neurological disease. He maintains an adjunct faculty appointment at the University. Dr. Nicholas’s post-doctoral studies were conducted at UCSF. His pre-doctoral work at both UCSF and Stanford University investigated germ cell development from both primordial germline and pluripotent stem cells. He received his Bachelor’s degree from the University of California, Berkeley. Prior to his interest in stem cell and developmental biology, Dr. Nicholas was a member of the discovery research team at Sugen, Inc.
Michael O'Connor, PhD
Director, Molecular Medicine Research Group
Western Sydney University, Campbelltown
Exploring new avenues for cataract treatment using human pluripotent stem cells
Cataract, or vision loss due to clouding of the eye’s lens, is a large and costly international problem. Over 80 million people currently have low vision due to cataract. Cataracts can only be treated surgically, and while this restores vision the ability to focus between near and far objects is lost. Cataract surgery is the most commonly performed ophthalmic procedure and globally costs tens of billions of dollars annually. Due to population aging the incidence of cataract is increasing, and so are the associated costs. It has been estimated that a 10-year delay in cataract formation could halve the number and costs of cataract surgery. To address this issue Dr O’Connor has developed a world-first method to produce large numbers of human lens cells from pluripotent stem cells. The ability to access unlimited numbers of human lens cells, for defining cataract risk factors and performing anti-cataract drug screening, represents a true paradigm shift in international cataract research.
Michael uses human pluripotent cells to model macular degeneration and cataract. He received his PhD from the University of Sydney in 2005, regenerating and characterising functional ocular lenses in vitro. During postdoctoral studies in Vancouver, Canada, he identified genes related to pluripotency and played a key role in the commercial development of mTeSR1 and TeSR2 with Stem Cell Technologies. As the current President of the Australasian Society for Stem Cell Research he has developed a touring stem cell art exhibition, seen by over 1 million people, that stimulates public discussion on emerging stem cell therapies
Hideyuki Okano, PhD
Professor, Department of Physiology
Keio University School of Medicine
Challenge toward Clinical trial for Spinal Cord Injury using iPS cell
In our previous preclinical studies, when neural stem progenitor cells (NS/PCs)-derived from hiPSCs were transplanted into mouse or non-human primate spinal cord injury (SCI) models, long-term restoration of motor function was induced without tumorigenicity, by selecting suitable hiPSCs-lines (Nori et al., 2011; Okano et al., 2013; Okano and Yamanaka, 2014). However, NS/PCs derived from certain iPSC-lines gave rise to late-onset tumorigenicity after transplantation (Tsuji et al., 2010; Nori et al., 2015). Here, to preclude these risks before clinical application, we developed molecular characterization of hiPSCs and hiPSC-derived NS/PCs together with transplantation to injured spinal cord of immune-deficient mice (Nori et al., 2015; Sugai et al., 2016). We investigated global methylation status of tumorigenic hiPSC-NS/PCs and found that aberrant hypermethylation of a tumor suppressor gene was induced along the passage. For addressing the safety issue, remnant immature cells or tumor-initiating cells should be removed or induced into more mature cell types to avoid adverse effects of hiPSC-NS/PC transplantation. Because Notch signaling plays a role in maintaining NS/PCs, we evaluated the effects of γ-secretase inhibitor (GSI) and found that pretreating hiPSC-NS/PCs with GSI promoted neuronal differentiation and maturation in vitro, and GSI pretreatment also reduced the overgrowth of transplanted hiPSC-NS/PCs and inhibited the deterioration of motor function in vivo (Okubo et al., 2016). Based on these findings, we are establishing methods of production, selection and transplantation of clinical grade NS/PCs stocks-derived from human iPSC stocks generated from HLA-homozygous super-donors by CiRA. We aim to commence clinical research (Phase I–IIa) trials for treatments of sub-acute phase SCI using hiPSCs-derived NS/PCs in the near future.
Professor Hideyuki Okano is Dean of Keio University School of Medicine and Team Leader of Laboratory for Marmoset Neural Architecture, Brain Science Institute RIKEN. In addition, Dr. Okano holds important posts including, University of New South Wales Visiting Professor (From 2009), University of Queensland Honorary Professor in the Queenland Brain Institute (From 2008), and Professor of Department of Physiology, Keio University School of Medicine (From 2001). In the past he served as Professor of Osaka University Graduate School of Medicine (Department of Neuroscience, 1997-2001) and Professor of University of Tsukuba (Department of Molecular Neurobiology, Institute of Basic Medical Sciences, 1994-1997). He also researched at University Tokyo and The Johns Hopkins University School of Medicine as instructor. Dr. Okano has been the recipient of numerous awards, most recently including the Molecular Brain Award (2016), Balz Award (2014) and receipt of the Medal with Purple Ribbon (From Japanese Emperor, 2009). His scientific research area is basic neuroscience, stem cell, and regenerative medicine including iPSC, NSC, and clinical application for spinal cord injury.
Alice Pébay, PhD
Associate Professor, ARC Future Fellow
University of Melbourne
Optimizing retinal cell differentiation of human pluripotent stem cells for large-scale disease modeling.
Human induced pluripotent stem cells (iPSCs) are valuable cells for retinal disease modeling, as these cells are of patient origin and can be differentiated into cell types of interest. This presentation will discuss the differentiation of iPSCs into retinal pigment epithelium cells and retinal ganglion cells and their molecular manipulations for modeling degenerative diseases of the retina and optic nerve.
Associate Professor Pébay is an Australian Research Council Future Fellow. She obtained her PhD in Neurosciences from the University of Paris VI in 2001 and subsequently joined Professor Martin Pera at Monash University to undertake research on human embryonic stem cells (hESCs). She then continued her research in this area at the University of Melbourne where she commenced in 2007. Since 2012, Associate Professor Pébay has been appointed to both the Centre for Eye Research Australia and The University of Melbourne. More specifically, Associate Professor Pébay heads research projects on the identification of signaling mechanisms involved in the maintenance of pluripotency, neural differentiation and more recently cardiac and ocular differentiation of pluripotent stem cells and diseased iPSCs. Her long-term research goal is the establishment of well characterized and efficient protocols for maintenance and differentiation of pluripotent stem cells, suitable for drug screening.
Martin Pera, PhD
Professor, Chair of Stem Cell Sciences, Program Leader, Stem Cells Australia
The University of Melbourne
Endoderm Progenitors in Health and Disease
Using human pluripotent stem cells as a screening platform, we have identified a novel cell surface maker that identifies foregut endoderm progenitors in pancreas and liver. We have shown that cells of the biliary reaction, a regenerative response to liver damage, premalignant and pancreatic ductal adenocarcinoma cells, and the columnar epithelium of Barrett’s Oesophagus, a preneoplastic precursor of oesophageal carcinoma, all express this antigen on their surface. The antigen is expressed in liver and pancreas during development, and is also found in the serum of patients with pancreatic or liver cancer. Cell surface antigens expressed on fetal stem or progenitors cells provide biomarkers of tissue regenerative processes, and can be used to monitor progression neoplasia if repair processes fail to resolve the underlying pathology.
Martin Pera is Professor of Stem Cell Sciences at the University of Melbourne, the Florey Neuroscience Institute, and the Walter and Eliza Hall Institute for Medical Research. He serves as Program Leader for Stem Cells Australia, the Australian Research Council Special Research Initiative in Stem Cell Sciences. His research interests include the cell biology of human pluripotent stem cells, early human development, and germ cell tumours. Pera was among a small number of researchers who pioneered the isolation and characterisation of pluripotent stem cells from human germ cell tumours of the testis, work that provided an important framework for the development of human embryonic stem cells. His laboratory at Monash University was the second in the world to isolate embryonic stem cells from the human blastocyst, and the first to describe their differentiation into somatic cells in vitro. He has provided extensive advice to state, national and international regulatory authorities on the scientific background to human embryonic stem cell research.
Karim Si-Tayeb, PhD
l'institut du thorax
From urine to the study of metabolic disease – A patient-driven strategy to decipher PCSK9 roles and functions.
In the last 10 years, PCSK9 emerged as a promising target for the treatment of autosomal dominant hypercholesterolemia (ADH). With the emergence of induced pluripotent stem (hiPS) cells and following differentiation protocols as a model for human pathology studies, we recently demonstrated the modeling of ADH due to mutations in the LDLR gene. Our strategy to better understand the role of PCSK9 included reprogramming somatic cells from patients carrying the S127R mutation into hiPS cells, differentiate these cells into hepatocytes and characterize the intracellular impact of the mutated form of PCSK9. To facilitate our strategy, we developed the isolation, culture, amplification and reprogramming of progenitor cells derived from urine samples (UCell). After reprogramming the hiPS cells (UhiPS), showed similar characteristics to hiPS cells classically reprogrammed from skin fibroblast. Through differentiation of the UhiPS cells hepatocytes were obtained that secrete PCSK9, internalized LDL particles and respond positively to statin treatments. To validate our model, in addition to generating hiPS cells carrying the PCSK9-GOF S127R, we reprogrammed cells from a patient carrying the PCSK9-LOF R104C/V114A and showed a spontaneously low level of LDL-cholesterol. Compared to control cells, UhiPS-S127R and –R104C/V114A differentiated in a comparable manner toward hepatocytes. In our hands, while hepatocytes carrying the S127R mutation showed a lower ability to uptake LDL, hepatocytes carrying the R104C/V114A mutations displayed the inverse, when compared to control cells. In addition, cells derived from the hypercholesterolemic patient responded positively to statin treatments, at a level comparable to the clinical response of patients carrying the same S127R mutation. Altogether, we demonstrated that differentiated hiPS cells are a relevant model to decipher PCSK9 functions and patient-derived urine samples represent a convenient source of somatic cells for such study.
After achieving his Ph.D. in Bordeaux under the direction of Dr. Jean Rosenbaum, Dr. Si-Tayeb joined the research team of Prof Stephen Duncan at the Medical College of Wisconsin, Milwaukee, WI. Together, they developed a strategy in order to instruct patient-derived hiPS cells to become liver cells and study metabolic diseases, including familial hypercholesterolemia. Back in France, and after a short passage in Paris, Dr. Si-Tayeb joined the “institut du thorax” in Nantes in order to work more specifically on PCSK9 together with Prof Bertrand Cariou, a M.D.-Ph.D. specialized in endocrinology who participated in clinical trials on PCSK9 inhibition and Dr Cédric Le May specialized in lipid metabolism. In their resource article published in Disease Models and Mechanisms, Dr Si-Tayeb and his team showed how it is possible to isolate cells from urine samples, amplify them, reprogram them into hiPS cells and then instruct them to become liver cells. Starting with urine samples of patients from the Nantes area, they showed that it was possible to model the effects of PCSK9 mutations on LDL uptake in a petri dish.
Bernie Tuch, PhD
Director, NSW Stem Cell Network
Honorary Professor, School of Medicine Sciences, Discipline Physiology,The University of Sydney
Unproven Interventions with Stem Cells
Autologous cell based interventions, which include mesenchymal stem cells, are mostly unproven therapies increasingly being applied for musculoskeletal and other medical disorders by medical practitioners, with limited regulatory control.
A self-regulated Code of Practice written for such Australian practitioners was released in February 2015. It required external review of the medical practices amongst other checks and balances that included (a) practising evidence based medicine; (b) ensuring fully informed consent was obtained from patients; (c) manufacturing the autologous product to be injected into the patient using internationally accepted standards; and (d) following the advertising standards set for medical practitioners by the Australian Health Practitioner Regulation Agency (AHPRA). Whilst the Code of Practice is beginning to be followed by some, with clinical trials approved by a Human Research Ethics Committee, and a Register of Adverse Events created, it may be too little too late. In July 2016, the NSW Coroner released findings of a case whereby an elderly person given autologous stromal cells derived from lipoaspirate to try and assist progressive dementia died within 24 hours of the procedure. This appears to be the first recorded death in Australia of a recipient of an autologous (stem) cell intervention. In September, the Therapeutic Goods Administration released a Consultation paper inviting feedback on proposed tightening of the regulations. These include no direct marketing to the public, upgrading of manufacturing standards, registering with the TGA, and reporting of serious adverse events.
If the regulations are indeed tightened, as now seems likely, the number of unproven autologous cell based interventions should diminish, as will the risk to recipients. However, at the same time innovation is likely to be reduced, as the cost for carrying out the interventions increases to meet the new standards required.
Dr Bernie Tuch is the Director of the NSW Stem Cell Network and a Professor of Medicine at both The University of Sydney and Monash Universities. He is involved in cutting edge research with clinical application, looking at novel ways of achieving positive outcomes. Attempts to examine the safety and efficacy of novel interventions have been through phase 1b/2a clinical trials, including the use of human fetal pancreas, and more recently encapsulated human islets, as a therapy for type 1 diabetes. His former research group was responsible for the creation of the first human embryonic stem cell lines at an Australian public hospital. Over the past few years he and colleagues have grappled with the issue of trying to move the stem cell field forwards expeditiously in humans with interventions, many of which are unproven.
Hans-Dieter Volk, PhD
Chairman, Institute for Med. Immunology & Berlin-Brandenburg Center for Regenerative Therapies
(BCRT) & Dept. Immunology, Labor Berlin Vivantes Charité GmbH
Immunomodulation and immunogenicity of human MSC-like cells: What did we learn from in vitro and in vivo studies?
Mesenchymal stromal cell (MSC) therapy is a promising option to support endogenous regeneration and immunomodulation. However, the clinical results are contradictory. We think that the recent studies have to major limitations: poor characterization of MSC(like) cell products which were used and the lack of adequate immune monitoring to better understand therapy response, mode-of-action, dose-dependency etc. Because of their high regenerative and immunomodulatory potency shown in various preclinical models and their well-defined manufactoring process in 3-D bioreactors, we focused on the characterization of placental-expanded mesenchymal like adherent cells (PLX) that are aimed to be applied as allogeneic off-the-shelf product.Interestingly, by minor manipulation in the manufactoring process, Pluristem generated two related products PLX-PAD and PLX-RAD. Both PLX-cell types were comparable regarding their in vitro differentiation capacity and marker profile (CD73+90+105+45-31-34-) that is typical for MSC-like cells. However, the cells showed different properties in some but not all preclinical models. We hypothesized that the protective effects are mediated by the PLX-cells´ secretome which might be different between PLX-RAD and PLX-PAD cells. In fact, conditioned medium of PLX-RAD expressed a distinct secretome and showed distinct effects in vitro and in vivo. Whole genome DNA methylation analysis and CD screen revealed significant differences between the two products.PLX-PAD cells are supportive in different tissue regeneration models. In fact, clinical phase I/IIa studies in severe chronic limb ischemia (CLI) and muscle injury demonstrated safety but also clear hints for efficacy. Immune monitoring gave insights into immunogenicity, immune modulation, and dose-related effects which help to design ongoing studies.In summary, extensive biomarker studies provided mechanistic insights into PLX products and highlighted the need for careful characterization of MSC-like cell products to better understand dosing, indication, mode-of-action etc.
Hans-Dieter Volk, MD, is Professor of Immunology and head of the both Institute of Medical Immunology, Charité Berlin and Berlin-Brandenburg Center for Regenerative Therapies (BCRT) as well as deputy spokesman of the Berlin-Brandenburg School for Regenerative Therapies (BSRT) (all Berlin, Germany). In addition, he is scientific head of the Division Immunology of the Labor Berlin Charité Vivantes GmbH, Berlin. His focus lies on implementation of new concepts in diagnosis and therapy of immunological diseases. Hans-Dieter Volk is an expert in coordinating and conducting clinical trials by biomarker development, monitoring new cell therapies, performing proof-of-concept and investigator-initiated trials (all phase I/II). Moreover he was/ is co-editor/editorial board member of several high-impact journals (e.g. Am J Transpl, Transplantation) and board member of several scientific medicine societies (e.g. German Society Immunology, German Society Sepsis).
Dustin Wakeman, PhD
Senior Research Scientist, Regenerative Medicine
Translating Pluripotent Stem Cell Therapies For Focal Brain Disorders
A major challenge for the clinical application of pluripotent stem cell therapy for neurodegenerative diseases is large-scale manufacturing and cryopreservation of neurons and glia that can be prepared for surgery with minimal manipulation. To address this obstacle, midbrain dopamine (iPSC-mDA) and forebrain (iPSC-FB) lineage neurons were derived from human induced pluripotent stem cells and cryopreserved in large production lots for biochemical and transplantation studies. Cryopreserved, post-mitotic neurons retained high-viability with gene, protein, and electrophysiological signatures consistent with that of the neuronal lineage. To test therapeutic efficacy, cryopreserved iPSC-mDA neurons were transplanted without sub-culturing into the 6-OHDA-lesioned rat and MPTP-lesioned nonhuman-primate models of PD. Grafted neurons retained midbrain lineage with extensive innervation of both rodent and monkey brain with no aberrant growth. Behavioral assessment in parkinsonian rats demonstrated significant reversal in functional deficits up to 6-months post-transplantation. In addition, cryopreserved iPSC-FB neurons grafted into the striatum of athymic NUDE rats survived and innervated distant anterior and posterior brain structures at 9-months post-grafting. These findings demonstrate a simple and efficacious surgical intervention to deliver cryopreserved iPSC-derived neurons for brain disorders and support translational development of pluripotent cell-based therapies in neurodegenerative disease.
Dr. Wakeman’s primary research goals are directed at determining the long-term value of stem cell based therapeutics for regenerative medicine. His past work using dopamine neurons derived from pluripotent stem cells, both human embryonic stem cells and induced pluripotent stem cells (iPSC), as a cell based strategy for dopamine replacement in animal models of Parkinson’s disease has consistently supported therapeutic value moving toward the clinic. Dr. Wakeman recently joined RxGen, Inc., a translational therapeutics and disease modeling company, where he is applying his expertise and experience in regenerative medicine to bridge the translational research gap using primate models of human disease. Dr. Wakeman also holds an Adjunct Assistant Professor position in the Department of Psychiatry at Yale School of Medicine.
For Research Use Only. Not for use in diagnostic procedures.