I used DMEM with 10%fbs and 1%ps for my complete media. I thawed the cells on monday now its been almost five days. The colour of the media is still red.Should i just let it grow?
It usually takes 24-48 hours for any cell line to divide. If its more than 3-4 days, then most probably the cells have achieved stationary phase. It might be due to the cells been passaged too many times and they have reached their limit. Try trypsinizing the cells and re-suspending them. Wait for 24-36 hours. If the same continues, then its time to discard them. Sometimes it might happen due to the FBS you are using. Due to improper storage condition of FBS, multiple freeze/thaw cycles of FBS leads to FBS degradation. This degraded FBS wont help in the growth/multiplication of your cells.
Hello Zikry Izzuddim,
Did you take the viability count after thawing? The cells look few and far apart. These cells may take a bit longer to get confluent.
I would suggest that if you have extra frozen vials of these cells, thaw two vials of the same passage and take the viability count. If the viability is too low, pool the cells from the two vials of the same passage and plate the cells in a culture vessel of smaller surface area so that the cells come closer to one another. The cells are happy when they are together, and revival is much faster after thawing.
You may leave the cells which are in culture aside, as they will take a while to get confluent. Do not trypsinize them at this stage because they are already under stress due to the thawing process. Meanwhile you may thaw new vial.
I feel the low viability count after thawing may be either due to improper freezing method or storage conditions.
Best.
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