FACS detection of myoblasts and fibroblasts

Peter Ashby 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 
surface markers.
> 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.


Peter Ashby
Wellcome Trust Biocentre
University of Dundee
Dundee, Scotland
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