We had recently re-organized the cell lines in our cryo can (Liquid nitrogen container) by shifting all the canisters into a new can. This process took about 10-15 minutes. Now, all the cell lines are revived and growing well. But two of our hERG expressing cell lines look like they have lost the expression. But are growing normally. Do you think this transferring procedure caused loss of gene expression?
This would depend on the entire procedure you use for cryopreservation. See for example: http://www.sciencedirect.com/science/article/pii/S001122401630178X
Hi Jaison. There is no universal explanation on how cells behave after being frozen and thawed.
I have seen some frozen cells become inviable after ~5 min exposure to RT and some cells that survive repeated failure of keeping LN2 supplied.
hello see this link
http://www.sciencedirect.com/science/article/pii/S001122401630178X
Thank you all for the answers. I read the paper linked above. However it was not very convincing because the exposure time I come across is between 5-15 minutes. It is quite obvious that Me2SO is hazardous to cells at such extended exposures at 37'C. Apart from this paper I find nothing. A good topic to work on I suppose.
Next time, put the frozen cells in different cryo cans, and avoid the transferring procedure. Technics related to tissue culture are not mathematical sciences.
Good luck.
- Badges
- Science method