Can anyone suggest why the D283 medulloblastoma cell line did not attach after thawing? What changes can be made to the dish surface or culture medium to solve this?
Hello Sergei Smirnov ,
It is difficult to be sure since you did not give much information about your thawing step : was it a tube that you bought from a bank, that was sent from another lab or that was frozen in-house ? If it is a new cell line (ie the 2 first options), I would not be too surprised by the poor quality of the frozen cells ... It could have absolutely nothing to do with the medium you are using, nor the culture surface. In this case, and if the unattached cells are still alive, just give them some time, maybe supplement your medium with more FCS (for example 20% instead of 10%), as it might help the cells to attach, but some cells don't like that so this is something I would try only if nothing else works.
On the other hand, if it is a tube that you froze yourself and a cell line that you are used to cultivating, maybe some changes in the medium might explain their difficulty to attach. Once again, if the cells are alive but not adhering to the surface, maybe just give them some time to get better after the freezing - thawing steps : after all, it is a stress for them.
Hope this helps a bit ! Good luck with your cells.
Dear Benedicte,
Many thanks for your answer! It is a tube from ATCC. 30% of cells are alive. But are not atteching.
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