1) I really stuck quite suddenly with my experiments because of strange issue with C2C12, upon differentiation the C2C12 (myofibers) layer become shrink-ed to one point and totally de-attach from remaining space in well, It never happened before just like condensed in one point and all other place in well is quite clear. I would be grateful if any experienced researcher with C2C12 can help me to sort out the issue.
During proliferation cells are OK even at initial stage of differentiation but later it happening.
Initially I though my cells are not healthy I changed to fresh cells, media everything except than same company 12-24 well plates, I dont know is it because of plate surface coating error or the differentiation media, usually I prefer to use fresh one, now I am thinking to use just freshly heat-inactivated horse serum, please help me to find and sort out the issue.
Or anyone can give some tip for seeding to reduce such chance
2)Any-one working with same cell line please share the protocol how to in-activate horse serum manually (instead of commercial one)
Thank you
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