[Histonet] cell characterization labels/stain

[Y. Wang]

I have a question about cell characterization. I know this isn't exactly 
your normal histology question but I've learnt so much from the people on 
this list in the past I was hoping someone would have a suggestion.

We have started a primary cell culture from porcine anterior cruciate 
ligament (ACL). We have so far harvested the ACL, minced it and placed the 
pieces in culture to let the cells grow out of the pieces. After 5 days or 
so we have cell out growth from the pieces, however, we have cells with a 
variety of cell morphologies. I always thought we could generally tell 
cell type from the cell morphology however, it is proving to be rather 
difficult as we have a range of 'spindle-like' cells from very 
spindle-like to sort of spindle like but a bit more triangular.

So, my question is: Does anyone know of a label/stain that can be used to 
characterize (ideally ACL) fibroblasts so that we can prove that our cells 
are ligament fibroblasts and not something else.

I have done a search in some cell culture books, on the web and of past 
papers. I've even found images of what people think are ligament 
fibroblasts growing out of ligament pieces (ACL and MCL) however, I can't 
seem to find any information on characterizing these cells with something 
more reliable than saying 'these are fibroblasts because they have the 
characteristic spindle-like morphology'(am I not looking in the right 
places or with the correct key words?). The closest I've come was a paper 
on primary culture of urethra fibroblasts, which they identified as being 
fibroblasts because they were not smooth muscle cells (labeled for SM 
actin) or endothelial cells (labeled with an endothelial cell marker)!

Any suggestions would be greatly appreciated.
Yak-Nam Wang

Senior Fellow
Department of Bioengineering
University of Washington
Box 357962
Seattle, WA 98195

Tel.: (206)-221-5873
Fax.: (206)-221-5874