Human Plasma-Like Medium (HPLM)

Create physiologically relevant cell culture models with HPLM

 

Gibco Human Plasma-like Medium (HPLM) is formulated to resemble the natural cellular environment found in the body, mimicking the metabolic profile of human plasma. 

 

Widely used, synthetic cell culture media, including MEM, DMEM, RPMI 1640, and DMEM/F-12 contain glucose, amino acids, vitamins, and salts at concentrations that, in large part, do not reflect those found in human plasma. These media also lack additional plasma components needed to mimic the metabolic profile of human plasma. When studying cancer and other diseases, results with more physiological relevance will enable researchers to help improve their understanding of human function and illness.

The HPLM solution

Gibco HPLM contains the same salt concentrations found in human plasma, as well as the same concentrations of over 60 polar metabolites, such as amino acids, nucleic acids, sugars, and small organic acids. In resembling the natural cellular environment found in the body, HPLM helps provide researchers the ability to study the impact of physiologically relevant cell media on their specific applications.

 

HPLM supplemented with fetal bovine serum (FBS) can support cell growth and viability comparable to those of conventional FBS-supplemented basal media formulations. For most cell lines, adaptation is not required to transition from conventional medium to HPLM.

 

HPLM is beneficial to your cell culture experiments in several ways:

  • Physiologically relevant—formulated with more than 60 polar metabolites and salt concentrations that resemble the natural cellular environment found in the body
  • Peer reviewed—extensive research publications completed using HPLM formulation
  • Easy to use—direct replacement for your current media when supplemented with FBS

Meet the inventor of HPLM

The inventor of human plasma-like medium is Jason R. Cantor.

 

As a postdoc at the Whitehead Institute/MIT in Cambridge, Jason set out to create what would become human plasma-like medium (HPLM), a physiologic medium designed to more closely reflect the metabolic composition of human blood, thus permitting the study of cultured cells in biochemical conditions with greater relevance to human physiology.

 

Cantor reported his development and initial studies using HPLM in early 2017 (Cell). Read his publication here: Physiologic Medium Rewires Cellular Metabolism and Reveals Uric Acid as an Endogenous Inhibitor of UMP Synthase.

We are proud to work with Jason to bring this innovation to market, and excited by the immense possibilities that HPLM could bring across diverse areas of the scientific community. As Jason notes, "The recent development of physiologic media, like other efforts designed to address the modeling capacity of cell culture, holds immense potential to improve understanding and interpretation of diverse biological and pharmacological studies." Read more from his 2019 commentary here: The Rise of Physiologic Media .

 

Jason is listed as an inventor on a patent application for HPLM assigned to Whitehead Institute.

Research results using HPLM

Research has shown that cellular performance is impacted by the use of HPLM, indicating that physiologic media can help increase the relevance of results from physiological studies.

 

HPLM rewires cellular metabolism

Diagram of the effects of different uric acid concentrations on uridine monophosphate synthase in humans and in mice

Graphical abstract summary: “Among the most prominent was an inhibition of de novo pyrimidine synthesis—an effect traced to uric acid, which is 10-fold higher in the blood of humans than of mice and other non-primates. We find that uric acid directly inhibits uridine monophosphate synthase (UMPS) and consequently reduces the sensitivity of cancer cells to the chemotherapeutic agent 5-fluorouracil. Thus, media that better recapitulates the composition of human plasma reveals unforeseen metabolic wiring and regulation, suggesting that HPLM should be of broad utility.”

 

Reproduced with permission from: Cantor JR, Abu-Remaileh M, Kanarkek N et al. (2017) Physiologic medium rewires cellular metabolism and reveals uric acid as an endogenous inhibitor of UMP synthase. Cell 169: 258–272.E17. doi: 10.1016/j.cell.2017.03.023

HPLM improves T lymphocyte activation

Diagram of the effects of different uric acid concentrations on uridine monophosphate synthase in humans and in mice

Graphical abstract summary: “Among the most prominent was an inhibition of de novo pyrimidine synthesis—an effect traced to uric acid, which is 10-fold higher in the blood of humans than of mice and other non-primates. We find that uric acid directly inhibits uridine monophosphate synthase (UMPS) and consequently reduces the sensitivity of cancer cells to the chemotherapeutic agent 5-fluorouracil. Thus, media that better recapitulates the composition of human plasma reveals unforeseen metabolic wiring and regulation, suggesting that HPLM should be of broad utility.”

 

Reproduced with permission from: Cantor JR, Abu-Remaileh M, Kanarkek N et al. (2017) Physiologic medium rewires cellular metabolism and reveals uric acid as an endogenous inhibitor of UMP synthase. Cell 169: 258–272.E17. doi: 10.1016/j.cell.2017.03.023

The functional characteristics of cells, including morphology and growth, in HPLM are comparable to those in conventional basal media formulations.

 

Human plasma-like medium maintains cell morphology

Figure 1. Gibco HPLM supports MCF7 cell culture. MCF7 breast adenocarcinoma cells were cultured in DMEM (Cat. No. 10566016) or Human Plasma-Like Medium (HPLM, Cat. No. A4899101) supplemented with 10% FBS (Cat. No. A3840101).

Figure 2. Gibco HPLM supports HeLa cell culture. HeLa human cervical adenocarcinoma cells were cultured in DMEM (Cat. No. 11965092) or Human Plasma-Like Medium (HPLM, Cat. No. A4899101) supplemented with 10% FBS (Cat. No. A3840101).

Figure 3. Gibco HPLM supports LNCaP cell culture. LNCaP human metastatic prostate carcinoma cells were cultured in RPMI 1640 (left; Cat. No. 61870036) or HPLM (right; Cat. No. A4899101) supplemented with 10% FBS (Cat. No. A3840101).

Human plasma-like medium maintains cell growth

Figure 4. Gibco HPLM supports comparable growth rates in continuous culture. Tumor cell lines MCF7, HeLa, A549, and THP-1 were grown in DMEM (blue) or HPLM (red) supplemented with 10% FBS (Cat. No. A3840101) for five passages. Cell number was assessed at the end of each passage and used to calculate the average population doubling time for each culture.

Human plasma-like medium supports cell growth in multiple cell types

Table 1. Cell types successfully grown in human plasma-like medium
Cell type tested
Cell origin
A375, adherent Malignant melanoma
A549, adherent Lung carcinoma
HCT116, adherent Colorectal carcinoma
HeLa, adherent Cervical adenocarcinoma
Jurkat, suspension T cell leukemia
LNCaP, adherent Metastatic prostate carcinoma
MCF-7, adherent Metastatic breast cancer
MDA-MB-231, adherent Metastatic breast cancer
NK, primary, suspension Lymphocyte, blood
NOMO-1, suspension Monoblastic/monocytic leukemia
PLB-985, suspension Myeloid leukemia
Sp2, suspension Mouse B lymphocyte
THP-1, suspension Monocytic leukemia
U-2 OS, adherent Osteosarcoma
WM115, adherent Malignant melanoma

The following customer stories feature the work of our Gibco HPLM product testers

“We used the HPLM media with cancer cell lines from different tissues and different mutation background. Our overall impression of HPLM is quite positive. HPLM media was suitable for all the tested cancer cell lines. Cells were easily and quickly adapted to the HPLM without affecting viability and cell culture performance. However, we detected relevant impact of cell proliferation, metabolism and mitochondrial function in cells grown in HPLM as compared with classical media. After our experience using HPLM in cancer cell models we consider it as the best choice to get more physiological data.”

Omar Torres-Quesada

PhD, Postdoctoral Researcher at University of Innsbruck

Austria

“Immunometabolism is an exciting area of scientific investigation that has immense potential for the development of new therapeutics. It is becoming clear that how immune cells rewire their metabolism after activation depends on what nutrients are available and in what quantities. Since we study metabolite transport in primary human immune cells, we find HPLM media to be essential for modeling metabolic flux in the most physiologically relevant way possible. We have extensively vetted the use of HPLM across CD4 T helper cell lineages and find that it supports both proliferation and effector function."

Justin Rettenmaier,

Associate Director at Jnana Therapeutics

Boston, MA

"Standard cell culture media, including the Dulbecco’s Modified Eagle Medium (DMEM), contain a non-physiological excess of nutrients, including glucose and glutamine. At the same time, they are low on uric acid. Nutrient availability, however, is a most critical factor for the regulation of mTOR calling for a careful consideration of culture conditions. Therefore, we employed Human Plasma Like Medium (HPLM) to gain a better understanding of mTORC1 regulation under physiological nutrient abundance. In line with mTOR’s intricate connection to nutrient sensing pathways, mTORC1 activity, as measured by Thr389 phosphorylation of S6K, was lower when U2OS cells were cultured in HPLM instead of DMEM. In HPLM media and irrespective of any treatment, AKT and the glucose-sensing AMPK were activated. Most importantly, however, we found that p53 was required to sustain low mTORC1 activity under HPLM culture conditions irrespective of Nutlin-3a treatment. Together, our investigation using HPLM revealed a nutrient-dependent role of p53 in mTORC1 inhibition."

Martin Fischer

Principal Investigator at Leibniz Institute on Aging

Germany

Webinar: Physiologic medium to study human cell biology

 

Environmental factors influence human cell physiology and can also affect drug efficacy, but existing model systems used to study human cells have limitations for understanding these contributions. In this webinar, Dr. Jason Cantor will discuss the initial development and use of human plasma-like medium (HPLM), a physiologic medium designed to more closely reflect the metabolic composition of human blood. By examining human cancer cell lines in HPLM versus traditional media, Dr. Cantor and colleagues have recently shown that HPLM has widespread effects on metabolism and gene essentiality, and further, that HPLM can be used to reveal new insights into metabolic regulation and drug efficacy.

 

Speaker: Jason R. Cantor, Investigator, Morgridge Institute for Research, Assistant Professor of Biochemistry, University of Wisconsin-Madison

Webinar highlights:

  • Complete media, the workhorses of cell culture studies, typically consist of a basal medium that poorly resembles conditions encountered in the human body and a largely undefined serum supplement
  • Human Plasma-Like Medium (HPLM) is a relatively new physiologic basal medium designed to more closely reflect the metabolic composition of human blood
  • The development of physiologic media, like other efforts designed to address the modeling capacity of cell culture, holds immense potential to improve understanding and interpretation of biological and pharmacological studies

Assessment of cancer spheroid formation in Human Plasma-Like Medium

 

Human Plasma-Like Medium (HPLM) is a cell growth formulation designed to mimic the metabolic profile of human plasma to help maintain the physiological state of cells.

We demonstrate the capability of HPLM to support the growth of cancer spheroids in several well-studied cancer cell lines

Figure 5. HPLM supports 3D spheroid formation. Representative images of spheroids from various cell types cultured in RPMI standard medium and Human Plasma-Like Medium (HPLM) that were grown for five days. Images were captured using the Invitrogen EVOS M7000 Imaging System. Scale bar = 650 µm.

Recent publications using HPLM

Title Cell type(s) Research area(s) Assay type
CRISPR screens in physiologic medium reveal conditionally essential genes in human cells K562, MOLM-13, SUDHL4, and NOMO1 Cancer biology CRISPR, RNA-seq, metabolite profiling, enzyme activity
De novo pyrimidine synthesis is a targetable vulnerability in IDH mutant glioma BT054 oligodendroglioma cells Cancer biology transduction, drug screening, metabolite analysis, mass spec, isotope tracing, western blotting
Early reduction of glucose consumption is a biomarker of kinase inhibitor efficacy which can be reversed with GLUT1 overexpression in lung cancer cells PC9, H1229, H3122 Cancer biology growth kinetics, drug screening, xenograft, transduction
Activating mTOR mutations are detrimental in nutrient-poor conditions  primary mouse embryonic fibroblasts Cancer biology growth kinetics
Therapeutic efficacy of RAS inhibitor trametinib using a juvenile myelomonocytic leukemia patient-derived xenograft model  juvenile myelomonocytic leukemia (JMML) cells Immuno-oncology western blot
Title Cell type(s) Research area(s) Assay type
Human Plasma-like Medium improves T lymphocyte activation human and mouse T cells Immunology transcriptome analysis, NGS
MTHFD2 is a metabolic checkpoint controlling effector and regulatory T cell fate and function primary human T cells Immunology T cell activation, metabolomics, mass spec, next-gen bisulfite sequencing, IHC, flow cytometry
Single-cell profiling of the antigen-specific response to BNT162b2 SARS-CoV-2 RNA vaccine human PBMC Immunology LIBRA-seq, single-cell RNA-seq, flow cytometry
Plasmacytoid dendritic cell activation is dependent on coordinated expression of distinct amino acid transporters plasmacytoid dendritic cells isolated from PBMCs Immunology ELISA, flow cytometry, isotope tracing, single-cell RNA-seq, CHIP-seq, IHC
mTOR regulation of metabolism limits LPS-induced monocyte inflammatory and procoagulant responses human PBMC Immunology ELISA, flow cytometry, metabolite profiling, isotope labelling, RNA-seq, ChIP
Obesity and inflammation influence pharmacokinetic profiles of
PEG-based nanoparticles		 Sk-Hep1 human liver adenocarcinoma cells, Tp1 cells  Immunology, Obesity fluorescence imaging, flow cytometry
Suppression of CEBPδ recovers exhaustion in anti-metastatic immune cells mouse NK cells Immunology siRNA mediated knockdown, tumoricidal assay
Title Cell type(s) Research area(s) Assay type
Physiologic medium rewires cellular metabolism and reveals uric acid as an endogenous inhibitor of UMP synthase K562, KMS12BM, NOMO1, P12-Ichikawa, SEM, SUDHL4, 786-0, A549, MCF7, SW620, BJ, CLF-PED-015T, primary acute myeloid leukemia cells Cancer metabolism growth kinetics, metabolite quantification and analysis, cholesterol quantification, oxygen consumption
Inhibiting both proline biosynthesis and lipogenesis synergistically suppresses tumor growth HeLa, MDA-MB-231, MCF-7, A549, HepG2, 8133, SKOV3, and mouse 4T1 cells Cancer metabolism electron production in metabolism, metabolite isotope tracing, RNA-seq
ZBTB1 regulates asparagine synthesis and leukemia cell response to L-asparaginase CUTLL1, SUPT1, MOLM-13 Cancer metabolism metabolite profiling, isotope tracing, ATAC-seq, mass spec
Lineage-specific silencing of PSAT1 induces serine auxotrophy and sensitivity to dietary serine starvation in luminal breast tumors  HCC1806, SUM149, BT549, HCC1937, HCC70, BT20, MCF7, MDA-MB-453, ZR75-1, EFM19, HCC1500, T47D Cancer metabolism metabolite analysis, cell proliferation, mass spec
Mitochondrial NADP+ is essential for proline biosynthesis during cell growth HEK293E, HeLa, K562 Cancer metabolism CRISPR, mitochondria isolation, spheroid growth, cell cycle, metabolic flux analysis, mass spec
Metabolic perturbations sensitize triple-negative breast cancers to apoptosis induced by BH3 mimetics  HCC1143, HCC1937, MDA-MB-231, MDA-MB-468 Cancer metabolism metabolite analysis, BH3 profiling, qPCR
p53-mediated AKT and mTOR inhibition requires RFX7 and DDIT4 and depends on nutrient abundance  U2OS, HCT116 Cancer metabolism  transfection, ChIP, western blotting
Increased mitochondrial proline metabolism sustains proliferation and survival of colorectal cancer cells RKO Cancer metabolism  transfection, cell proliferation, apoptosis, mass spec
Human Plasma-Like Media fine tune mitochondrial function and alter drug sensitivity in cancer cell lines SW620, MCF7, A375 Cancer metabolism cell proliferation, respirometry
Human acute leukemia utilizes branched-chain amino acid catabolism to maintain stemness through regulating PRC2 function human primary AML and ALL cells Cancer metabolism metabolome analysis, flow cytometry, transplantation, RNA microarray, ChIP-seq, cell cycle, isotope tracing
Methionine metabolism controls the B cell EBV epigenome and viral latency P3HR-1 Burkitt’s lymphoma cell Cancer metabolism CRISPR, MeDIP, ChIP, RNA-seq, mouse xenograft, metabolite profiling, IHC
Elevated transferrin receptor impairs T cell metabolism and function in systemic lupus erythematosus mouse naïve T cells, patient-derived T cells Cancer metabolism  CRISPR, RNA-seq, mass spec, IHC
Formate overflow drives toxic folate trapping in MTHFD1 inhibited cancer cells SW620 (CLL-227), HCT116 (CLL-247), MDA-MB-468 (HTB-132) and MDA-MB-231 (HTB-26) cells Cancer metabolism drug affinity responsive target stability (DARTS) assay, cellular thermal shift assay (CETSA), stable isotope tracing, mass spec
Canagliflozin impairs T cell effector function via metabolic suppression in autoimmunity human PBMC-derived T cells T cell metabolism ELISA, flow cytometry, metabolic analysis by extracellular flux analyzer, western blot, stable isotope tracer analysis (SITA) by LC-MS, RNA-seq, proteomic analysis, mass spec
Iron is critical for mucosal-associated invariant T cell metabolism and effector functions human PBMCs T cell metabolism transferrin uptake assay
Targeting fatty acid synthase in preclinical models of TNBC brain metastases synergizes with SN-38 and impairs invasion MDA-MB-231 Cancer metabolism drug screening, RNA expression
Hypoxanthine in the microenvironment can enable thiopurine resistance in acute lymphoblastic leukemia NALM-6, REH, SEM, CEM, Jurkat Cancer metabolism drug assay (viability readouts using Cell-Titer Glo, Caspase 3/7 assay)
Conditional lethality profiling reveals anticancer mechanisms of action and drug-nutrient interactions K562, SEM, NOMO1, P12-Ichikawa Cancer metabolism drug screening
Glucose limitation protects cancer cells from apoptosis induced by pyrimidine restriction and replication inhibition Jurkat, Raji, A549, HCT116, MDA-MB-468, HEK293FT Cancer metabolism apoptosis
Coordination between the eIF2 kinase GCN2 and p53 signaling supports purine metabolism and the progression of prostate cancer Human prostate cancer tumoroids (TM00298) Cancer metabolism cell cycle, metabolomics, ribosome biogenesis, CRISPR screening
Title Cell type(s) Research area(s) Assay type
Nickels and tines: the myth of nickel allergy in intracranial stents not applicable Medicine/surgery nickel allergy test
Generation of induced pluripotent stem cell-derived beta-cells in blood amino acids-like medium human iPSCs  Stem cell differentiation qPCR
The development of a smart magnetic resonance imaging and chemical exchange saturation transfer contrast agent for the imaging of sulfatase activity NA (proprietary ligand linked to gadolinium) Imaging enzyme activity fast field cycling NMR relaxometry, z-spectrum
Quantification of cell death and proliferation of patient-derived ovarian cancer organoids through 3D imaging and image analysis patient-derived ovarian cancer organoids Drug screening high content imaging and analysis
A new physiological medium uncovers biochemical and cellular alterations in Lesch-Nyhan disease fibroblasts primary human skin fibroblasts from healthy individuals and those with Lesch-Nyhan disease Disease biology scratch assay, qPCR, mitochondrial respiration
Vitamin A resolves lineage plasticity to orchestrate stem cell lineage choices mouse hair follicle stem cells Stem cell differentiation ATAC-seq, RNA-seq, CUT&RUN (CNR)
Human plasma-like medium (HPLM) induces Cryptococcus neoformans in vivo cell morphologies Cryptococcus neoformans (C. neoformans)  Host-pathogen interaction flow cytometry
CRISPR-based functional profiling of the Toxoplasma gondii genome during acute murine infection toxoplasma gondii Host-pathogen interaction genome wide gene survey
Engineered myovascular tissues for studies of endothelial/satellite cell interactions Primary human myogenic cells, primary Human endothelial progenitor cells Tissue engineering cell-cell interaction, cell differentiation

Order Gibco HPLM

Item
Cat. No
2-hydroxybutyric acid (available through Alfa Aesar) Alfa Aesar A18636-03
Fetal bovine serum, dialyzed, US origin 26400044
6-Well Plate, TC Surface, Pack of 1< 140675
96-Well Plate, TC Surface, Pack of 1 167008
150 mm EasYDish, TC Surface, Pack of 10 150468
DPBS, no calcium, no magnesium 14190144
TrypLE Express Enzyme (1X), no phenol red 12604013
Trypsin-EDTA, 0.05%, phenol red 25300054
Trypan blue solution, 0.4% 15250061
Countess 3 Automated Cell Counter AMQAX2000

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