FACS detection of myoblasts and fibroblasts
p.r.ashby at dundee.MAPS.ac.uk
Fri Oct 6 10:50:00 EST 2000
In article <200010061222.OAA04537 at intwww.zit.med.uni-tuebingen.de>,
Wolfgang.Schechinger at med.uni-tuebingen.de ("Wolfgang Schechinger")
> Hi all,
> I'd like to characterize by FACS analysis a myblast cell culture
> derived from a human skeletal muscle biopsy. There probably grow
> some fibroblasts, so I have two main cell populations.
> As far as I am aware there is no myoblast specific marker, thus I'd
> do the following:
> 1) stain almost all cells with anti MHC-1
> 2) stain fibroblasts with anti-Thy.1 (CD-90)
that sounds like the best way, I'm not aware of any myoblast specific
> 1a) stain myoblasts with anti NCAM, since I assume that fibroblasts
> will not express this surace antigen
I don't remember the details but NCAM expression in developing muscle is
very dynamic and don't forget that if these cells are from a biopsy you
will have satellite cells and fibroblasts rather than embryonic
myoblasts and the two are not the same.
I think you will be better off labelling the fibroblasts and do a
negative on the muscle cells. There is an old method to enrich for
myoblasts by plating the cells for a short time then taking of the
unattached cells and repeating the procedure with them. It works by
exploiting the fact that fibroblasts are stickier than myoblasts.
Wellcome Trust Biocentre
University of Dundee
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