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Attention-seeking cell cultures - reduce subculture ratio?


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#1 seanspotatobusiness

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Posted 19 October 2010 - 08:06 AM

The cells I'm culturing (QM7) need to be passaged at least every two days, sometimes even every day. The ATCC recommends a subculturing ratio of 1:4 to 1:5 but what's this for? Optimal expansion perhaps? What if I passage them 1:10; then I could get on with my life in-between passaging! I will confirm with my supervisor before making this change, but would like input from the forum so I'm better prepared.

Edited by seanspotatobusiness, 19 October 2010 - 08:06 AM.


#2 rkay447

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Posted 19 October 2010 - 10:03 AM

Some cells require contact in order to grow and if you passage them too thin they will go into senescence or die but ATCC indicates that allowing these cells to go to confluency is a bigger problem. Next time you split your cells, why not make one dish at 1:5 and another at 1:10. This way, if the 1:10 stops growing, you still have the 1:5 going and you haven't lost anything. Another possibility is to use a lower FBS level. Although most people use the 10%, this is for maximum growth. I will culture cells with 3-5% FBS if I want cells to grow slower.

#3 seanspotatobusiness

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Posted 21 October 2010 - 01:59 AM

Some cells require contact in order to grow and if you passage them too thin they will go into senescence or die but ATCC indicates that allowing these cells to go to confluency is a bigger problem. Next time you split your cells, why not make one dish at 1:5 and another at 1:10. This way, if the 1:10 stops growing, you still have the 1:5 going and you haven't lost anything. Another possibility is to use a lower FBS level. Although most people use the 10%, this is for maximum growth. I will culture cells with 3-5% FBS if I want cells to grow slower.


Hey. Thanks for the advice. I spoke to my supervisor and he agrees re: reducing the ratio. Since we lower the FBS level as part of the protocol for differentiating them however, he doesn't want to do that. I currently lack the knowledge/experience to say whether reducing the FBS would have any additional effects which might adversely affect our experiments later on so I'll have to do as he says for now. :)




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