[Histonet] cell characterization labels/stain
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.
Department of Bioengineering
University of Washington
Seattle, WA 98195
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