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3T3-L1 differenciation and maintenance

adipocyte differenc cell culture insulin 3t3-l1 3t3

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#1 fredUNH

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Posted 08 February 2012 - 03:12 AM

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

#2 bob1

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Posted 08 February 2012 - 03:51 PM

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

#3 fredUNH

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Posted 09 February 2012 - 12:07 AM

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.

#4 AdipoFat

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Posted 09 February 2012 - 05:19 AM

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

#5 fredUNH

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Posted 09 February 2012 - 06:45 AM

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





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