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how to culture cancer stem cells? - help (Jul/15/2008 )

wacko.gif
hi, guys! I am going to do some research about cancer stem cells, does someone else work in the same field?
here I have a question that how to culture putative cancer stem cells? should I culture them as normal stem cells? or ....

can someone provide a protocol for me to follow?

thanks in advance! wub.gif

-karyomit-

QUOTE (karyomit @ Jul 16 2008, 12:59 PM)
wacko.gif
hi, guys! I am going to do some research about cancer stem cells, does someone else work in the same field?
here I have a question that how to culture putative cancer stem cells? should I culture them as normal stem cells? or ....

can someone provide a protocol for me to follow?

thanks in advance! wub.gif


I'm doing the same work...if you are using primary cells from patients, please use collagenase I buffer to digest the tissue. After homogenisation, cell sorting is followed, CD133 and CD34/44 are used. After sorting, please use Stem cell medium to culture them... the formulation for 100ml stem cell medium is 90.8ml DMEM/F12-GlutaMax, 2ml STEMPRO┬« hESC SFM Growth Supplement (50X), 7.2ml BSA 25%, 80ul FGF-basic (10 μg/mL), 2ml P/S (100X). Make the medium freshly. I just add 150ul (for 24 wells plate) and 500ul (for 6 wells plate) stem cell medium every two days, and seldom change medium until the cells are full in the well since there are sphere formation.

-Leonard-