FACS detection of myoblasts and fibroblasts

[Peter Ashby]

In article <200010061222.OAA04537 at intwww.zit.med.uni-tuebingen.de>, 
Wolfgang.Schechinger at med.uni-tuebingen.de ("Wolfgang Schechinger") 
wrote:

> 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

-- 
Peter Ashby
Wellcome Trust Biocentre
University of Dundee
Dundee, Scotland
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