First of all cells will be cultured on monolayer and than will be transformed to suspension. What is a suitable culture medium for each phase? Does anyone have any protocol or tips to help me with this method?
We use Ham's F12::
http://www.lifetechnologies.com/order/catalog/product/21700075
The 10 X1 L should come as 10 individual packets of powdered medium. It's also available as prepared medium. We don't usually grow ours in suspension, but it shouldn't matter.
You might check if the cells are from ATCC or DSMZ or other cell collectors. If so, you could find detailed culture protocols on their websites.
Dear Terry,
Thanks for tour answer. I'll try it. Do you use only FBS as supplement?
Dear Rongqing,
I appreciate your answer but I forgot to say that I'm trying to find an alternative culture medium.
Yes, %10 FBS. Ham's F12 was specifically developed for growth of CHO-K1, however Rongqing has a good point. Those are good resources to check. We also use Normocin to protect against Mycoplasma contamination.
http://www.invivogen.com/normocin
In FBS 10% and DMEM F-12 HAm available with sigma. the cHO cell grow very well and after monolayer u can make the suspension by using the EDTA and trpsin solution..
even u can check the medium invitrogen or other available locally
Hello
CHO cells should be cultured in Ham’s F12K . In addition, you can try with DMEM-12 modified with 10% FBS, RPMI-1640, IMDM with 10% FBS , and alpa-MEM.
Good Luck
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