I am using TC-100 media(with serum and antibiotics) having sodium bicarbonate and L-Glutamine to grow Sf21 cells, but after I thaw them, they initially attach and grow for some time, then they fail to attach after 2-3 passages and eventually die.
What could be the possible reason for it? How can this be tackled?
Hi, it is possible that you have subcultured badly Or in the passage process, a trypsinization shock is applied to your cells
and the cells generally are damaged by something that you may done .
Considering your query, there might be 2 chances.
1. Probably you have revived the old stock or you have preserved them in -80 for a pretty long time. (usually, the cells which are revived from liquid nitrogen are always better than the cells revived from -80).
2. trypsinization shock. Usually, if the trypsinization time is prolonged, the cells lose their capacity to adhere and cannot gain morphology.
I recommend you revive the fresh stock and culture again.
I will like to suggest this manual page 16-17. It should assist you.
https://tools.thermofisher.com/content/sfs/manuals/Insect_Cell_Lines_UG.pdf
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