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Eagles09's Content

There have been 4 items by Eagles09 (Search limited from 22-August 18)


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#43254 Subculture in low glucose media

Posted by Eagles09 on 07 November 2009 - 11:01 PM in Tissue and Cell Culture

Isra, yes this will slow their growth rate, as they are losing their preferred growth substrate. Are you culturing them in low glucose long-term or short-term? I only have experience culturing them in low glucose short-term, and I grow my cells in regular (high glucose) media until they are just shy of their ideal confluence (so, my ideal confluence is 80%, and I grow them to about 70-75%), and then I add low glucose media for 12 or 24 hours. So yes, it is better to do it the way you are currently employing.



#43212 Stimulating/Drugging Mammilian Cells

Posted by Eagles09 on 07 November 2009 - 12:45 PM in Tissue and Cell Culture

Thank you for the responses. I did perform an experimental screen diluting the stock concentration of drug into a larger volume of media and then distributing, and this seemed to be effective, as you are both confirming. So, I will stick with this method. Agreed about the pipetting inaccuracies, but until robotics is standard (and therefore more available), there isn't a good way to get around it! I always think to myself how in 30 years from now I will teach a class and jokingly tell my students how lucky they are because we used to have to pipette manually :D



#42773 How to seed the cells evenly in 6 wells plate

Posted by Eagles09 on 05 November 2009 - 07:05 AM in Tissue and Cell Culture

Thana, which techniques did you end up using? I seem to be having the same problem and it is throwing off my numbers.



#42770 Stimulating/Drugging Mammilian Cells

Posted by Eagles09 on 05 November 2009 - 07:01 AM in Tissue and Cell Culture

Does anybody know the proper way to stimulate a mammalian cell culture with drugs?
I am using well plates, and am unsure if I should add the drug directly to each well containing medium (e.g., a 1:100 dilution of pre-diluted drug in DMSO to DMEM), or I have also read a proper way to do it is make a stock solution of drug in the medium using a 1:1000 drug:DMEM dilution.
If anyone has experience, please help. These are kinase inhibitors.




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