In the last decade, organoid cultures have quickly become a popular way to create “mini-organs” to support the advancements in the study of organogenesis, disease modeling and subsequently the development of new therapies. Scientists are creating lab-grown miniature versions of organs that so far include kidney, liver, brain, prostate and pancreas, that more closely resemble the composition and functionality of organs.
There are many protocols, tools and techniques that can be used for organoid cultures – ranging from microplates, extracellular matrices, hydrogels, and bioprinting to microfluidics. Depending on your cell type, research area and ultimate goals, the options can seem overwhelming.
So how does one know where to start? Learning from the work that has already been done is a great place to start. We’ve assembled a team of Corning Life Science experts, who are ready to answer your organoid-related questions. Corning has over 30 years of experience in 3D cell culture and offers some of the original and most widely used 3D tools, such as Corning® Matrigel® matrix, Transwell® permeable supports and the Corning spheroid microplate.
Corning experts include Feng Li, Senior Scientist Development, Hilary Sherman, Applications Scientist, Himabindu Nandivada, Senior Development Scientist and Nitin Kulkarni, Sr. Scientific Support Specialist. Dr. Feng Li has been working on 3D hepatic model systems for liver toxicity and disease modeling. His recent work includes establishment hepatic 3D spheroid culture procedures, testing primary human hepatocytes (PHHs) for 3D spheroid culture, and assay development for chronic liver toxicity testing and repeated-dosing with PHH spheroids in Corning ultra-low attachment spheroid microplates. Hilary Sherman is an Applications Scientist with Corning Life Sciences. She has worked with a wide variety of cell types including mammalian, insect, primary and stem cells in a vast array of applications, including 3D cultures. Dr. Nandivada has more than 10 years of experience in human pluripotent stem cell culture and material science. Dr. Kulkarni has worked in Scientific Support group for several years supporting 3D cell culture and recently presented talks and a webinar on surfaces used for organoid culturing.
0 0 0 Total: 0 A spheroid can be a mere aggregate of one or more cell types where cells do not attach to a surface other than to other cells to form a spheroid. Ultra low attachment spheroid microplates are an ideal tool to form and assay spheroids in the same microplate. Organoids, on … Continued
0 0 0 Total: 0 Corning® Matrigel matrix is one of the most successful animal based extracellular matrix that has been used to culture organoids. There are multiple reports on engineering a synthetic matrix to grow organoids with the studies being compared to their morphology/functionality in Matrigel matrix. Researchers have succeeded in generating them for … Continued
Is there an efficient way to make organoids compatible with a high throughput drug screening environment?
0 0 0 Total: 0 High-throughput screening with organoids is an emerging technology and scientists are looking for efficient ways to address it. Corning® Matrigel® matrix is the matrix of choice for organoid workflows and lends itself well to the growth of various types of organoids. There are two commonly used methods to dispense organoids … Continued
0 0 0 Total: 0 Organoids can either be recovered from the hydrogel/extracellular matrix or they can be stained in the matrix itself depending on what matrix and concentration is used. For recovering organoids from Corning Matrigel matrix, the culture can be treated with Corning Cell Recovery Solution (not Dispase as it will make single … Continued
I feel one of the biggest challenges in organoid culture is in delivering nutrients and gas exchange especially as the organoids grow. Thoughts or recommendations?
0 0 0 Total: 0 Organoids need a continuous feed of fresh nutrients and waste removal as the culture expands in size and since they are not vascularized, as suggested in your question. There are 3 recommendations that have been used for this purpose: Researchers have been successfully culturing and maintaining organoids embedded in Corning … Continued
What is the timeline for organoid culture? When should I expect to begin to see the beginning of formation, timing for passaging, media changes?
0 0 0 Total: 0 Developmental progression of organoids and the timing of different steps in organoid culture are dependent on origin of cells, protocol being used (e.g. media formulations) and the desired organoid type. In some protocols, organoid-like structures or buds can be observed as early as 24 hours after organoid culture process is … Continued
What are your recommendations for culture vessels that work best with organoids? Something that works well with imaging too.
0 0 0 Total: 0 The type of culture vessels used for organoid culture is dependent on the protocol being used for organoids. Different culture vessels are used at the various stages of generation, culture and characterization of organoids. Tissue culture-treated vessels (multi-well plates or dishes) can be used together with a natural (Corning Matrigel … Continued
I am growing intestinal organoid from iPSC cells in Corning Matrigel matrix. I plan to collect the organoids from the matrix for subsequent tests and analysis. Can you please recommend a protocol for harvesting?
0 0 0 Total: 0 We recommend Corning® Cell Recovery Solution, 100mL (Product #354253) for harvesting organoids from Corning Matrigel matrix. Below is a general guideline, using an example of six week old organoids in 96-well spheroid microplates. Incubation time may be optimized for best results. Wash wells twice with 150 µL cold PBS (organoids … Continued
What do you think about the research that discusses organoid co-culture with endothelial cells? Do you find that the organoids can get adequate nutrients with good culture media?
0 0 0 Total: 0 At this point in vivo organoids do not have their own vasculature but endothelial cells have been shown to form rudimentary vascular structures in some organoids like liver buds. When they are implanted into immune-deficient mice, the host vasculature fuses with these structures to form functional vascular network and eventually … Continued
0 1 0 Total: 1 Spheroids and organoids are both 3D structures made of many cells. Although this terminology has been interchangeably used there are distinct differences between them. An organoid is a “collection of organ-specific cell types that develops from stem cells or organ progenitors and self-organizes through cell sorting and spatially restricted lineage … Continued
Are there any special media requirements for organoid culture or is it sufficient to just address the specific needs of that cell type?
0 0 0 Total: 0 It depends on the organoid of interest as described by a recent commentary by Sato and Clevers. Researchers have been developing and optimizing their favorite organoid medium for kidneys (Xia Y. et al. (2014) Nature Protocols. 9:2693) (Freedman B. et al. Nat Commun. 2015; 6: 8715), brain (Lancaster M.A. and … Continued
Is there any evidence of successfully using organoids as working as mini organs for drug testing? I know the thought is there but is anyone using this successfully?
0 0 0 Total: 0 There is growing interest in using organoids in drug screening and the pharmaceutical industry is now looking to use the platform as well. However, there are still many challenges to overcome to make this technology routine for drug-screening. The first successes were documented in a subset of cystic fibrosis patients … Continued