Hi everyone! I am having problem in growing Huh7.5 cell line. I am using cDMEM (invitrogen 11965) supplemented with non-essential amino acids (1X) and pen/strep. At beginning after thawing and seeding, cells attach to surface but later on they become detached and lead to death. I am trying again and again but having same problem. Please help me!!
Nasir,
Huh7.5 cell line are temperature and growth sensitive, as soon as you take them from the 37 C incubator or if the cells are very confluent, they detach from the surface. I would recommend, you should coat your dishes/plates with Poly-L-Lysine and then grow Huh7.5 cells on them.
You could find the protocol here
http://www.sigmaaldrich.com/technical-documents/articles/biofiles/poly-lysine-product.html
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