JOHN B
Posts: 1
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Joined: 25/4/2006
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HUMAN PRE-ADIPOCYTES
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Dear colleagues,
Currently, we are trying to isolate pre-adipocytes from human omental tissue.
Culturing pre-adipocytes in a commercial system (ZenBio) seems to work very well, but costs a lot of money.
For that reason and aiming to isolate (pre)adipocytes from fat distributed at different body parts, we would like to develop our own isolation procedure for (pre)adipocytes.
We now perform a collagenase I digestion on omental tissue. This seems to work because we do get cells (pre)adipocytes but there are also other cells attaching to the culture surfaces resulting in overgrowth of our target cells . Most probably mesothelial cells (cytokeratin stain +, factor VIII -). We also tried to incubate the digest in a (red cell) lysisbuffer, but this didn't make much of a difference.
So here is my question: can anyone provide with a pre-adipocyte isolation protocol and/or give any ideas about how to get rid of the contaminating cells.
Thanks for all your suggestions
JohN
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