There are many types of Matrigel Matrix available to coat plates for cell culturing. Many of them are specifically designed to culture specific cell lines (for example specific for stem cell differentiation). In my current project I need to start culturing human pancreatic β cell line (EndoC-βH3 cells), which require pre-coating of plates with Matrigel and Fibronectin. I am little confused which matrigel matrix I should select? It would be great help if anyone can give me advice on this issue. Thanks.
I recommend
Matrigel (Sigma-Aldrich)-
Also you can read this article:
Aggregation of Engineered Human β-Cells into Pseudoislets: Insulin ...
doi ‹ https://journals.sagepub.com
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EndoC-βH3 monolayer cells were seeded onto Matrigel (Sigma-Aldrich)- and fibronectin (Sigma-Aldrich)-coated 96-well plates (TPP) at 7 × 104 cells/well, or groups of 50 day 3 or 6 PIs were handpicked up.
We use growth factor reduced Matrigel from Corning Life Sciences. This is especially good to use if you want to study the effects of growth factors on your cells.
Manash K Paul Heather Wanczyk Mohamed Elkablawy Thanks for your answers. I am planning to use both sigma and corning matrigel (as I managed to get both matrigels). I will share with you the results. I might need suggestions for further trouble shootings also.
Gabriel Liguori Thanks for the suggestion! I would also consider using MatriXpec along with other commercial ECMs to optimize the protocol. Thanks again.
The only matrigel I have had any luck with regarding endoc-BH1s is corning's hESC grade matrigel combined with fibronectin (bovine origin) and collagen Type-1 (rat origin). ECBH1s are notoriously difficult to grow and to get them to stay on the plate is even harder. I assume you will eventually end up performing a GSIS, in which case for them to stay on the plate is nothing short of a miracle when switching from 2.8/3G to 16.7G.
Also just saying we use 'corning matrigel' is highly unspecific. There are multiple different types of matrigel available depending on the cell type/morphology you wish to culture.
For these specific cells, we recommend using: https://ecatalog.corning.com/life-sciences/b2c/US/en/Surfaces/Extracellular-Matrices-ECMs/Corning%C2%AE-Matrigel%C2%AE-Matrix/p/354277 (corning cat# 354277) which is a matrigel qualified for hESC culture with mTeSR.
Also, I would like to point out that the Ravassard lab has its own formulation for the coating of dishes for ECBH1s, which is commercially available from the supplier for ECBH1s.
Thanks a lot Mirza Muhammad Fahd Qadir for sharing your experience. It would help a lot. I would update here my experiences with EndoBH1 culturing and functional analysis. I may need further help from you! Thanks again.
Abu Saleh Md Moin No worries. Always here to help. Good luck!
Mirza Muhammad Fahd Qadir Hope everything going well with you. had few queries on EndoC cell culture,
> While coating the plates with Matrigel+fibronectin+collagen, what sequence you followed for coating? I meant to say which one you used first to coat plates? Or you mix them all and coated? If you use one by one, how long for each coating? Is collagen absolutely necessary? I am asking in many papers they did not mention about collagen.
> According to the previous papers, The EndoC BH1 culture media should be serum free. However, in one paper I found these lines "Cells were passaged using TRYPLE 1x (Thermo Fisher, Waltham, MA, USA) and neutralised with heat inactivated human serum from human male AB plasma (Sigma Aldritch, Steinheim, Germany)." Can you suggest on this? Should we use human serum in the media during passaging of EndoC BH1 cells?
> If possible would you mind to share the protocol that worked for you in your lab.
Sorry for asking many questions! We received the only vial of Endo C cells and it took almost a year!
Thanks and kind regards
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