So i am trying to establish a cell line in our lab, and i asked myself: If i have the feeling on day 2 or 3 of the initial cell culture, that my cell density is too low, what options do i have? Is it better to wait and hope that the cells are not dying or is it possible to subculture them in this early state of culture, which would mean trypsinisation and centrifugation (so stress for the cells)?
what would you do?
thanks a lot!
Hello Anika Dreier
Early subculturing is not recommended. I presume you are working with primary cells. If I were you, I would better wait and hope for the best. It would not be possible to subculture them at an early state of culture because primary cells are very sensitive, and trypsin can be harmful to the cells and passaging primary cells too early or too frequently is not beneficial to them, and as you rightly mentioned, trypsinization and centrifugation would mean more stress to the cells who have just begin to recover.
Best.
I advice to subculture them in a smaller surface (for example in 4 wells of a 12 well plate instead of a T25 flask) to provide a better cell-cell contact which is a momentous factor for expansion of adherent cells.
thank you, you two for your answers. They are not primyry cells, it is a cell line but it is new to our lab...
so i did a subculture at about 50% confluency and it worked, the are growing fine now.. lets hope it stays this way :)
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