Protocol Online logo
Top : New Forum Archives (2009-): : Tissue and Cell Culture

3T3-L1 differenciation and maintenance - (Feb/08/2012 )

Dear all, I want to use 3t3L1 cells to work on differenciated adipocytes:
Different protocols are available:
I have some questions concerning different steps:
can I use FBS for proliferation instead of calf serum?
Is it necessary to use insulin in differenciated adipocyte maintenance medium (used 2 days after the insulin media and 4 days after differentiation initiation)?
Thank you very much.

Best regards,
Fred

-fredUNH-

FBS (foetal bovine serum)= foetal calf serum, just a name change to reflect the source, as other animals also have calves.

-bob1-

Thank you, but I think that serum from newborn calf is also available and used fir this cell model. I knew that calf and bovine was the same.

-fredUNH-

Hey, we normally use 10% FBS-supplemented DMEM all the way from undifferentiated 3t3-l1 fibroblasts to mature adipocytes. In the past we have also used DMEM-F12 supplemented with FBS or calf serum for preadipocytes. All of them work just fine.

We also use standard differentiation medium (IBMX, ins, dex) for 2 days, then another 2 days w/ insulin-supplemented medium and finally we go back to the 10% FBS/DMEM (no insulin).
We get cute and round adipocytes with this method.

www.adipofat.com

-AdipoFat-

Thank you very much AdipoFat, your message corresponds to what i generally see.!

-fredUNH-