Would anyone has experience culturing Huh7 cell line please help me. I first started to culture the Huh7.5.1 cell line about 3 months ago, and I noticed the cells started to grow on top of each other and I couldn't get confluent cells. So I started a new vial of cell that I stored in the -80 freezer. The first passage of the cell grew normal. However, in the second passage the cells did attach to the bottom of the flask, but had no obvious growth after 5 days. I need to figure out what is the problem, either the method I stored the cells or cultured the cells is not proper.
I cultured the cell with DMEM +10% FBS (Gbco)+1% P/S. I stored the cells by adding 10% FBS (Gbco)+10%DMSO without P/S. Any suggestions will be welcome and appreciated.
Hi Yanmei, I haven't cultured Huh7 cells but I have experience with HepG2 cells.
They are liver carcinomas and they grow clumpy. That is normal. They do not form a monolayer like epithelial cells.
Most cell lines should be stored in LN2 in 10% DMSO, 90% FBS. Storing cells at -80oC can be a problem. These cells have been acclimated to certain growing/freezing/thawing conditions.
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