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How many cell passages to be safe ?


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6 replies to this topic

#1 AnneC

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Posted 18 November 2004 - 10:16 AM

Hi !!

I don't have any experience in cell culture at all and I've just started to culture and transfect some cells for a few weeks. People from my lab gave me some SK-N-SH and C6 glioma cells.
The thing is nobody told me how many passages these cells have already been through... and this doesn't seem to worry anyone in the lab.
The cells look OK but I'm just wondering if they might lose some of their properties bit by bit with many passages...
What do you think ? Does anyone have experience with these cell lines especially ?
Thanks for your answers.

Anne

#2 Simonsays

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Posted 19 November 2004 - 05:55 AM

I've nerver worked with your cell lines, but what I personnaly do is working and 'pass' my cells for at least 40 passages, without a problem. I work with my cells as long as they "work well", and then I thaw some...
Hope this helped...

Simon

#3 Sprag

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Posted 19 November 2004 - 07:39 AM

some of the cell lines out there, such as HeLa, CHO, Cos-7 etc etc have been through countless number of passages. I would bring up fresh cells as soon as I see morphological changes, or if by mistake they've gone too confluent.

#4 DevGrp

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Posted 19 November 2004 - 08:15 AM

A couple of years ago I got an undergraduate student working on endothelial cell receptor expression to look at the changes in expression of 3 receptors over 25 passages of EAHY926 endothelial cells. The expression of ERbeta dropped by 300 fold, IcamI by 250 fold and GR by 8 fold (qRT-PCR). This seemed to explain why a previous student couldn't detect any ERbeta in her EAHY cells which had been high passage.
After this we always went back to the early passage stocks every 15 passages.
I think different cells behave in different ways. always keep an eye out for changes in the phenotype you are studying.

#5 postdoc

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Posted 19 November 2004 - 09:17 AM

I was wondering whether any phenotype changes have to be caused by genetic changes in the first place in high passage cells.

#6 sea_liu

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Posted 06 December 2004 - 07:39 AM

generally, some properties will change with the passages prolonged,

it is hard to say whether it is safe or not , because different cells have different situation cultured in vitro

#7 Jacen

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Posted 07 December 2004 - 03:32 PM

We always "bank" our cells. Meaning thaw a vial out, grow it up in culture, passage at least 3-4 times all the while expanding it to be as large as possible then freeze down the cell line as one batch. This is your stock that you grab from once you have used cells to a high level of passage. Every time you passage a cell line a selection is made and genetic drift from the original stock cell line will undoubtedly occur.




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