muscle vs. fat cells - (May/23/2007 )

Hi All,
I've been given the fun job of trying to culture primary intramuscular fat cells. However i've established that, with the size of the sample I can get, simply cutting the fat out is near impossible, so I thought I'd ask the question, does anyone know of a way of separating fat and muscle cells?

I want to culture the fat cells, treat them with all sorts of goodies in differentiation, then extract RNA and run RT-PCR.

Hi, I´ve been working alot with both primary muscle and fat cultures. Question is what kind of fat cultures you are going for... If you are interested in getting the mature fat cells then its a piece of cake, just take your cell suspension, centrifuge at 700 g and you´ll se the skeletal muscle cells pellet and the fat cells will float on the surface. If you want the preadipocytes for doing real cultures, then you need to centrifuge as before but remove the upper fat layer and carefully collect the infranatant without(!) disturbing the cell pellet!
After that, just check you cell cultures for skeletal muscle contamination using a desmin antibody, should work with antibodies for myogenic factors as well but desmin is quite reliable.

Hope this helps!

D