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How to primary culture adipocytes?


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#1 kuangzj

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Posted 23 April 2002 - 08:51 AM

Primary culture adipocytes: how to discard the oil floated on the surface?

#2 murusa

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Posted 10 September 2013 - 08:53 PM

I have the same problem, I isolated adipocytes and SVF (stromalvascular fraction) from mouse fat pads and there is huge volume of oil in cell cultures.. numerous washings helps to reduce it but despite this some volume of fat remains in culture.. as I know oil is toxic for cell culture..

does anyone know how to separate adipocytes from the oil released from disrupted mature adipocytes?



#3 bob1

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Posted 11 September 2013 - 03:32 PM

You should be able to pellet the cells to remove the oil.  Oil should float whereas cells will sink.



#4 murusa

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Posted 12 September 2013 - 10:41 PM

Thank you! I isolate two fractions from mouse adipose tissue - mature adipocytes and stromal-vascular fraction.

 

In the case of SVF - yes, it is true.. I just aspirate and waste supernatant and wash cells in a pellet a few times.. although even in this case oil remains covering the tube walls so I should change tubes after each washing to reduce oil. So for this culture this way works..

 

But everything is different in the case of culture of mature adipocytes. They are big rounded cells full of fat inside, so they don't sink after centrifugation, they are always floating above media and in the plate wells also.. and there is always some volume of oil around them because they are very fragile and might be disrupted by pipetting that results in additional oil release.






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