Passage-hepatic stem cells - (Mar/30/2005 )

Hi,

I'm pretty much new to cell culture so forgive me if this sounds like a lame question: How important is passage in say a 5 day period? I'm trying to culture hepatic stem cells... and haven't acyually done passage at all yet...

tanya

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Tanya,

Check out Dr. James Sherley's work over at MIT - supression of asymmetric cellular kinetics (SACK). If what you have are truly hepatic stem cells they will not replicate exponentially until you abrogate the effects of p53 by replacing the missing downstream product of IMPDH (p53 downregulates it) with something like xanthosine or hypoxathine. This will restore guanine nucleotide biosynthesis and lead to exponential proliferation.

Removal of xanthosine (or hypoxathine) will restore asymmetric cell kinetics and slow expansion.

Not a lame question at all - cutting edge, really.

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