Hello, I'm working with iPSCs and culturing them on feeder-free conditions using Matrigel. I'm using the Corning Matrigel for Organoid Culture as that was the only Matrigel that was being shipped in a reasonable amount of time. But now I'm having some trouble with cell attachment.
The colonies are rolling up after a few days. I'm not sure if it's because of the Organoid Matrigel or because of ROCKi removal (because I notice the detachment after I remove ROCKi as well) or a mix of both.
Has anyone faced this particular issue before or noted any differences in monolayer culture with the two different Matrigels? From what I can understand from the product documentation, both Matrigels are EHS tumor ECM with the only difference being the Organoid one being optimized for organoid culture. But the components are ultimately the same.
I've been growing spheroids in regular Corning Matrigel and have had no problems. I don't know what ROCKi is.
Corning® Matrigel® matrix for organoid culture (Cat. No. 356255) is a kind of growth factor reduced Matrigel matrix which is recommended for organoid culture. Matrigel matrix for organoid culture is pre-screened to support the organoid application and minimizes variability across different lots. It has been verified to support mouse intestinal organoid growth for more than seven passages with typical budding morphology and marker expression. It has also been verified to support polarized 3D structures of primary human airway epithelial cells expressing typical markers. Additionally, each lot is measured for its elastic modulus indicative of gel stiffness and is qualified to form stable “3D domes” commonly used in organoid culture protocols.
A normal Matrigel matrix, usually referred to as Matrigel basement membrane matrix (Cat. No. 354234), is not growth factor reduced which contains higher GF concentrations than Matrigel matrix for organoid culture. Matrigel basement membrane matrix is recommended in general cell culture and vessel coating.
Both products might vary lot-to-lot when being used in iPSC culture. The most stable performance in iPSC culture is brought by hESC-qualified Matrigel matrix (Cat. No. 354277). hESC-qualified Matrigel matrix is recommended for culture of hESC, stem cells, and iPSC.
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