Liquid nitrogen was finished, so I shifted all cells into -70°C for few hours. Is this correct? Does anything happens with the cells?
In my experience nothing should happen to your cells. I know labs where cells are abitually stored in a -80°C freezer without any problem. To be sure, I would defreeze and check a vial.
In my experience nothing should happen to your cells. I know labs where cells are abitually stored in a -80°C freezer without any problem. To be sure, I would defreeze and check a vial.
Yeah, I was thinking the same, that I should revive one vial to check whether cells are in good condition or not!!
Hi Pryianka,
The cells should be fine, we did this before. If most cells survive the temperature shift from liquid nitrogen to 37 degrees when seeding, then placing them to -80 will be fine.
Priyanka, for sure nothing will happen to the cells so long as they do not thaw; -70 C or -80 C does not permit thawing. Many times we have stored frozen cells in -70 to -80 C for months before transferring into liquid Nitrogen and from Liquid Nitrogen to the freezers without any notable negative effect. You should be fine with the cells.
@ priyanka viabilty will affect a bit less than -120 shows affect if u stores for longer period below to this temp
HI Priyanka, Dont worry, nothing will happen to your cells in -80C as long as you kept the vials frozen and did not allow them to thaw during transport. Hope you noticed the LN2 shortage in the tank relatively early and transferred the vials before any dramatic rise in the temperature and damage to the frozen cells. -70C should be fine for a short term storage but make sure you put tem back to LN2 tank if you want to keep a good quality, long term preservation of your cells.
Hey nothing should happen ideally. We had the same crisis in our lab too and we tried the same. We checked the revival also, which was pretty much the same on 3 cell-lines we checked. You may not worry, just check with your culture once you shift it back.
Good luck.!!
Hi, also in my experience I can say that no significative harm can come for this passage. You migth think of using a liquid medium to shift faster to -80 if you have to freeze them at -80 rigth away, usually isopentane (2-methyl butane) or acetone work fine.
Hello Priyanka , here in my lab we generally store our cell lines stock at -80 C only, we get healthy cells, no contamination issues. The only point of concern is , one should not allow the stocks to thaw while shifting.
@neha singhal -80 storage can protect the samples for few months only , if we wanna store for years liquid nitrogen is only the best option , samples stored in -80 for very long periods wont behave good in cultures rather they will die in multiple washing steps.
Hello everyone, as many pointed out -80°C is just fine for cell lines, also for long-term (most of these cell lines are tough ones). But: you should never put BACK to liquid N2 cells that have been shifted from N2 to -80°C. Just thaw them, let them recover and prepare new stocks. Cells just don't like up-and-down temp shifts. Hope that helps.
Raju Yadav- you are absolutely right, we do not store them for long. The stocks kept in -80 C are revived before two months. Though we are sometimes bound to use old stocks also, but in that case, it varied from condition to condition, sometimes the cells behaved unexpectedly and we have taken them till 20th passage, but sometimes they just die off.
@neha singhal exactly neha , I observed the same , i think it because of the way we collected blood and separation , as in our study patients will be Immune suppressants , so some times drug toxicity matters
Most frozen cells will survive -70 for a few months. After that, their viability will start to decrease, thats why LN2 is used for long term storage. So long as the vials did not thaw the cells should be pretty much OK.
Cells should be OK, but try to put them into liquid nitrogen as soon as you get a refill.
I do not know what was delay in between! There will be a certain percentage of loss associated with shifting of cells from liquid nitrogen to-70.Least delay-lesser percentage of loss.
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