Protocol Online logo
Top : Forum Archives: : Cell Biology

thawing of fridged cell culture - Reviving of cells (Jul/17/2006 )

Hi ,
any body could be explan why during reviving period of cells from liquid N2 needed hight density i a
little media and prone to contamination. Second things which way we know whether cells infected with mycoplasma .


A higher density would be needed due to the possibility of cell death during the processes of freezing or thawing. One reason they are prone to contamination is that when you thaw them often times you are thawing them in a waterbath and if you are not careful and submerge them too much or forget to clean off the vial with EtOH they are likely to become contaminated (waterbaths are not the cleanest of equipment ... though we try hard to keep them that way). Mycoplasma contamination is very hard to tell through just visual observation. There are several kits and tests you can use to determine mycoplasma contamination.

The topic on mycoplasma detection was covered before in the forum (link and link).


during cell revival, a lot are died and won't be interesting for you
tha'ts why a relative high density is recommended.
I think also that's the reaon of high density lies in the fact certain cell lines or cell types doesn't survive in loww density (IMR90 for ex or TrHBMEC... as i know these types)

Best way to know mycoplasma is to do a PCR on cell culture medium supernatant