Organoids, spheroids, and the study of cells as 3D models show great potential in many applications including disease modeling and regenerative medicine

Complex biology in a physiologically relevant context

Organoids, spheroids, and the study of cells as 3D models show great potential in many applications including disease modeling and regenerative medicine. 3D cellular models like organoids and spheroids offer an opportunity to better understand complex biology in a physiologically relevant context where 2D models have not proven as successful. Gain confidence to culture and analyze organoids and spheroids through validated protocols and educational resources brought together to enable 3D model success.

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Gibco Geltrex Matrix

Basement membrane, comparable to Matrigel, ideal for all types of cell culture applications including organoids

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EVOS FL Auto 2 Imaging System

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Research stories

Organoid research offers the possibility of starting regeneration in a dish, in a very high complexity because a 3D culture allows very high complexity structure and I think they hold huge promise for personalized medicine and therapy.

—Luigi Aloia, Research Associate, Gurdon Institute

Can you introduce yourself and your research?

I am Luigi Aloia. I work as a postdoc in the Laboratory of Maryhook at the Guldon Institute in Cambridge. I work on liver regeneration, and I use the 3D organoid structure as the main model of my research. Our goal is to improve the current therapy and the current approach to liver regeneration by understanding the basic molecular mechanisms behind it.

What excites you the most about organoid models?

Organoid research offers the possibility of starting regeneration in a dish in very high complexity, because a 3D culture allows very high complexity structure and I think they hold huge promise for personalized medicine and therapy.

Do you have any tips or tricks for anyone working with organoids?

The main tip is thinking in a three-dimensional way. When you work with 2D culture you seed your cell in plastic and then you see them growing in a monolayer. But with the organoids you can really follow the organization of the structure in a three dimensional way. This is an importance which—in your mind as a researcher—it gives you a more complex idea of how the tissue is formed, is maintained, and is repaired after injury.

What are the biggest advancements you have seen in this field?

I think in the last few years organoid cultures have improved a lot. So researchers are trying to optimize the culturing conditions and to obtain better performance of the culture in vitro and also in vivo when you transplant the culture, for instance, in animal models. And also there is a huge effort in the field now of understanding the molecular mechanism behind this culture system that will also help in understanding the molecular mechanism responsible for the tissue homeostasis and repair upon injury.

What labs have paved the way for you to expand?

Probably I consider the work in the Clever lab in the Netherlands. There are a bunch of endodermal-derived organoids from endodermal-derived tissues and organs, and this proves that both highly self-renewing organs and very slowly self-renewing organs such as the liver (which I work with) can give rise to organoid cultures, so we can study an organ in a dish. I also have to mention the Knoblich lab in Vienna where they establish these brain organoids, which is then followed by Madeline Lancaster here in Cambridge. So in this very moment, Cambridge is an important hub for organoid cultures.

What do you think is next for organoid research?

I think organoids hold a huge promise for personalized medicine and improving the current therapies in several diseases including cancer. I am convinced through the organoids we will learn more about the tissues, the homeostasis, and the injury, and the disease, and then we will be able to implement better therapies and better approaches.

I am really interested in how we can use organoids to model tumor pathology because so far we are only using 2d cell lines and animal models and now thanks to the organoid technology notably in prostate cancer, colon cancer and pancreas cancer, we have a better input in this pathology.

—Laura Broutier, Gurdon Institute

Can you introduce yourself and your research?

My name is Laura Broutier, and I am a postdoctoral fellow in Meritxell Huch’s lab at Gurdon Institute of Cambridge, and I am working on modeling primary liver cancer using organoid technology.

What is your primary goal in learning more about 3D/organoid technologies?

So far in the primary liver cancer field there is no good model to actually understand this pathology because 2D cell lines are failing to mimic the histoarchitecture of the primary liver cancer and the genetic landscape of this tumor. So using organoid technology we would like to try to get better model that could allow us to better understand these pathologues.

When did you first become interested in organoid research?

During my PhD, I was working with 2D cell lines and cancer cell lines, and I wanted to learn a bit more about cancer stem cells and stem cells in general. I thought that organoid technology would allow me to learn more about these stem cells and progenitor cells, so I think it’s a good model that could fill the gap between 2D cell lines and animal models, so I wanted to learn this technique as a skill for my future career.

Do you have any tips or tricks for anyone working with organoids?

When I started to use organoid models I wanted to do a large-throughput screening, and the common procedure you can find in the literature is seeding your cells in 24 multiwell plates. But when when you want to have a huge amount of cells, you can actually seed them in 6 well plates doing multidrop in the well and that will help you to limit the number of plates in the incubator.

What are the biggest advancements you have seen in the 3D cell culture field?

I am really interested in how we can use organoids to model tumor pathology because so far we are only using 2D cell lines and animal models. And now thanks to the organoid technology notably in prostate cancer, colon cancer and pancreas cancer, we have a better input in this pathology because we’re maintaining the cell-to-cell interaction, the cell-to-matrix interaction and that changed the answer to response of drugs.

What do you think is next?

I think the next step at least in my field (like modeling cancer using organoid technology) is really to go towards personalized medicine and large drug screening using this model. Now from one patient biopsy you can get one tumoroid line and you can use this line to test several drugs and then according to the response to this drug you can go back to the patient and you can treat this patient with appropriate drugs.

I think the most important advancements are being able to model disease and set up drug screening systems to have novel approaches and personalized medicines.

—Margherita Yayoi Turco, Royal Society Dorothy Hodgkin Fellow, University of Cambridge

Can you introduce yourself and your research?

My name is Margherita Turco. I am a Royal Society and Dorothy Hodgkin Research Fellow at the Center for Trophoblast Research, which is in the University of Cambridge. And I work on human reproduction, so I’m particularly interested in studying how the endometrium which is the lining of the uterus regenerates at every cycle and how it prepares for pregnancy, and this has been really difficult to study the human context because there have been no physiologically relevant or functional models. So I’ve been trying to set up organoid systems to model the human endometrium in vitro.

Do you have any tips or tricks for anyone working with organoids?

I think it’s very important to get to know your cultures really well, so how they grow, what they should look like. So you should monitor them well and passage them at the right time so that you have healthy optimally growing organoids for your experiments. What I found also really useful is using electronic pipetters for passaging the organoids, because it saves a lot of time and you can do it very reproducibly.

What do you think is next for organoids?

Organoids model the epithelial component of the tissue, so its lacking the endothelium for example and the immune cells and this can be an advantage because you are working with a clean system. But for what I am studying the stromal cells of the endometrium are very important and also regulating gland activity so I think the next challenge would be to be able to find robust co-culture systems to model the organoids - the epithelial component together with the stromal components to have a more complete model of the tissue.

What would you like to see adopted into the organoid workspace?

I think it would be important to have defined and tailored matrices in which to grow the organoids. So matrices that more accurately replicate the composition and the stiffness of the extracellular matrix of your tissue of interest I think would be great.

What are the biggest advancements you have seen in this field?

Well organoids have been now derived from almost all the major organs, and they have been really important in helping us understand about the tissue formation and the cell in each relationship. But besides that I think the most important advancements are coming in the field of really being able to model disease and especially setting up drug screening systems to have novel approaches and personalized medicines, and I think we are already starting to see that with the recent publications.

Key protocols and methods

There is a diverse range of protocols in literature for 3D culture – some that work, others that do not. Below are the most cited publications for each tissue type.

Brain

Liver

Intestine

Kidney

Lung

Brain

Liver

Intestine

Kidney

Lung

Culture and analysis systems enabling organoid, spheroid, and 3D cell models

Our culture and analysis systems are designed to enable the generation of organoid, spheroid, and 3D cell models.

Researchers utilize cell lines often to elucidate disease models of interest. Gibco cell lines allow you to closely mimic the in vivo state and generate more physiologically relevant data for organoid and spheroid research.

Custom cells

Primary liver cells

Stellate cells—non-quiescent cryopreserved myofibroblastic hepatic stellate cells (MF-HSC) isolated from adult human livers

LSECs—cryopreserved liver sinusoidal endothelial cells isolated from adult human livers

Stem cells

Human episomal Cas9 iPSC—A Cas9 expressing vector was stably integrated into Human episomal iPSC via lentiviral delivery and subsequent antibiotic selection

Contact us for assistance with your custom cell project

Selecting the right culture matrix is an important first step in developing a successful culture system for organoids and spheroids. Scaffold-based (or monolayer) systems are used to more easily transition from a 2D monolayer environment to 3D models. Scaffold-free (or suspension) systems are used for scale up and full organ development. Porous membrane-based systems are advantageous when polarization of the epithelial cells are needed in cell differentiation and tissue formation.

Gibco media and reagents are cited in organoid and spheroid peer reviewed papers are widely used in the growth, differentiation and maturation of cellular 3D models. This is where a culture system can be developed to suit specialized cell types like stem cells or cancer cell lines.

View all growth factors 

Pluripotent stem cells

Skin cells

Cancer spheroids

Hepatic cells

Brain cells

EVOS XL Core cell imaging system

Growing 3D models is a large investment in time and resources, and you need reassurance that your investment is going to give you the 3D models that you anticipate. Our portfolio of gene expression and visualization tools allows you to monitor the formation of your organoids and 3D models to help give you confidence that you are heading in the right direction.

Imaging instruments and analysis software

Confirming that 3D cell structures are maintaining the appropriate physiological morphology is paramount to reaching successful research outcomes. Our imaging and high-content analysis platforms and reagents have been recognized as trustworthy systems for analyzing organoid and spheroid cultures. In addition, Invitrogen antibodies are validated to ensure specificity and reproducibility in research results.

Researchers utilize cell lines often to elucidate disease models of interest. Gibco cell lines allow you to closely mimic the in vivo state and generate more physiologically relevant data for organoid and spheroid research.

Custom cells

Primary liver cells

Stellate cells—non-quiescent cryopreserved myofibroblastic hepatic stellate cells (MF-HSC) isolated from adult human livers

LSECs—cryopreserved liver sinusoidal endothelial cells isolated from adult human livers

Stem cells

Human episomal Cas9 iPSC—A Cas9 expressing vector was stably integrated into Human episomal iPSC via lentiviral delivery and subsequent antibiotic selection

Contact us for assistance with your custom cell project

Selecting the right culture matrix is an important first step in developing a successful culture system for organoids and spheroids. Scaffold-based (or monolayer) systems are used to more easily transition from a 2D monolayer environment to 3D models. Scaffold-free (or suspension) systems are used for scale up and full organ development. Porous membrane-based systems are advantageous when polarization of the epithelial cells are needed in cell differentiation and tissue formation.

Gibco media and reagents are cited in organoid and spheroid peer reviewed papers are widely used in the growth, differentiation and maturation of cellular 3D models. This is where a culture system can be developed to suit specialized cell types like stem cells or cancer cell lines.

View all growth factors 

Pluripotent stem cells

Skin cells

Cancer spheroids

Hepatic cells

Brain cells

EVOS XL Core cell imaging system

Growing 3D models is a large investment in time and resources, and you need reassurance that your investment is going to give you the 3D models that you anticipate. Our portfolio of gene expression and visualization tools allows you to monitor the formation of your organoids and 3D models to help give you confidence that you are heading in the right direction.

Imaging instruments and analysis software

Confirming that 3D cell structures are maintaining the appropriate physiological morphology is paramount to reaching successful research outcomes. Our imaging and high-content analysis platforms and reagents have been recognized as trustworthy systems for analyzing organoid and spheroid cultures. In addition, Invitrogen antibodies are validated to ensure specificity and reproducibility in research results.


Extend your organoid and 3D model research arm

Extend your organoid and 3D model research arm

Let’s face it, researchers today are in a difficult position. They are continuously being asked to transition their research into more physiologically relevant models, but the path to 3D models is not always obvious. Our cell model services will extend your impact by utilizing our dedicated team of cell biology experts to provide a custom 3D cell model solution.

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