how to purify the primary preadipocyte culture? - many fibroblast contamination (Oct/08/2008 )

hello, I dont know if there anyone is familiar with preadipocyte culture - so called adipose-derived stromal cells, from human liposuction samples.

I did try several times, but in the culture there are lots of fibroblasts. If anyone can suggest something in the protocol of culture, which could directly give a relatively purer preadipocyte culture?

thanks a lot

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Hi there,

I know of a group that use such cells to reconstruct 3D adipose tissue (PMID :17374391)

And the protocol they use to isolate preadipocyte is rather simple.

What makes you say that you have a lot of fibroblast in you culture?

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thanks a lot for your quick response. Actually for this moment, the supposed contamination of fibroblast is based on the morphology of cell culture. I attached our culture pics below. More they reach near confluency, they show typic fibroblast like looks, so some expeienced colleague told me they are fibroblast, I could just directly throw it in the trash bin.

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Have you tried to induce those cells to differentiate?

As you surely know, 3T3-L1 preadipocytes are fibroblast-like cells. So looking to these cells do not make me think of anything that would prevent me to try to induce their differentiation.

I may be wrong, though ^^

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Sorry for late coming. Thanks for your further suggestion.

I feel the same as you. Morphologically, some people say this cell is fibroblast-like cells, others say that they are anyway polygonal shape, and different from those bone marrow stromal cells. I also feel that it is difficult to judge simply by cell morphology. I already start to make CFU test and differentiate test. Hope it will give more information, I will learn from that.

thanks again, perhaps I will come back with those further results

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