Hi,
I want to isolate and culture macrophages from rats (different sites like bone marrow, peritoneal fluid etc.)
Most of the reported literature says to use the supernatant from mouse fibroblast cell line, L929 for differentiation of the macrophage culture (which I don't have unfortunately).
Does any one have experience using supernatant from other fibroblast cell line like mouse NIH 3T3 and human Hs27 cell line?
There may be less effect when using a different species like human but I am wondering if the above mentioned fibroblast cell line supernatant will have identical effect.
Thanks.
Debashish
I only have experience working with mouse/human cells but I guess the same principles apply. L929 cells secrete M-CSF and hence L929 conditioned medium is used as a substitute for recombinant M-CSF. Whether it would work on rat cells, I do not know. As to the questions of using alternate fibroblast cell lines, it would depend on whether these cells can secrete M-CSF and to what extent. If you end up using them, you may have trouble getting it published, as there may be no precedence. The best thing to do would be to get some recombinant rat M-CSF and use that.
Thanks, Julie. I am trying my best to get a vial of L929 cell line to complete my experiments.
I have experience of isolating peritoneal macrophages from mice. Macrophages can be cultured in RPMI-1640 medium.
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