Protocol Online logo
Top : Forum Archives: : Cell Biology

Slow cell growth when thawing cells - (Aug/07/2006 )

I grow CV1 cells. Whenever I thaw a new batch cells grow horribly slowly and need at least a week to become confluent. After this they grow ok and I split them every 2-3 days. This problem I have with different batches of cells, so it is not batch specific.
Anyone else had this problem?


I think it's quite common that cells grow slower right after thawing. They need few days to "rescue".


It's a very common phenomenon. So you needn't worry about it.


Agree with others.

It also depends on how much cells are in the vial u thawed. I usually split a confluent flask into 5 vials so it doesnt take that long to grow when I thaw them.


Hi ,
Its usual things because when we thawing a cells vials its take a time to regaining their normal metabolic activity and also needed certain chemokine for their normal growth .These all absent in the media because its secreated by same cells thats why its needed time to coming in normal growing and dividing stage of a cells after thawing.


thaw as fast as possible, dmso is toxic
with 293 I thaw and next day split them 1/8
for other cells I wait 2 or even 3 days


After thawing cells, plate them and change media in 2-3 hrs to get rid of the DMSO. Some people centrifuge it and wash it once with PBS to remove DMSO.


when i thaw them i put the contents of the vial into a t=25cm flask, passage this into a t=75cm flask the next day and then into a t=175 after that - works quite well for me. once skipped the middle size flask and they took forever to reach confluency . . . .