Although very basic, we need help: We have tried to isolate epithelial cells from cornea, however, we have in our culture dishes two different cell populations (morphologically speaking): epithelial-like and mesenchymal-like cells. What would be the best markers (for both populations) to distinguish them? We have in our lab: beta catenin, pan-cadherin and vimentin.
In general, epithelial cells express high levels of E Cadherin and very low levels of N-Cadherin and vimentin than mesenchymal cells. With reference to corneal cells, I would recommend you to go through this article
http://www.jbc.org/content/early/2013/07/10/jbc.M113.458141.full.pdf+html
I think you should try to use cytokeratin-3 marker to distinguish your corenal epithelium from MSCs, which (theoretically at least) should not have it.
In general, epithelial cells express high levels of E Cadherin and very low levels of N-Cadherin and vimentin than mesenchymal cells. With reference to corneal cells, I would recommend you to go through this article
http://www.jbc.org/content/early/2013/07/10/jbc.M113.458141.full.pdf+html
Thank you Zerina and Sridhar. I had seen papers using citokeratin-3 as a corneal epithelium marker, however, we dont have that antibody. I was wondering the feasibility of using beta catenin and pan-cadherin (I expect that epithelial cells would stain) and vimentin (I expect that mesenchymal/fibroblasts would stain). But what about: would mesenchymal/fibroblasts stain with beta catenin and pan cadherin?
Agree with Sridhar. Vimentin should be good. Not sure about other two.
Mesenchymal cells stain beta catenin relatively higher than epithelial cells
Not sure how this applies to corneal epithelium, but it seems like many types of epithelial cells become vimentin positive in vitro:
Kasper, M. and P. Stosiek, The expression of vimentin in epithelial cells from human nasal mucosa. Eur Arch Otorhinolaryngol, 1990. 248(1): p. 53-6.
Mendez, M.G., S. Kojima, and R.D. Goldman, Vimentin induces changes in cell shape, motility, and adhesion during the epithelial to mesenchymal transition. FASEB J, 2010. 24(6): p. 1838-51.
Rogel, M.R., et al., Vimentin is sufficient and required for wound repair and remodeling in alveolar epithelial cells. FASEB J, 2011. 25(11): p. 3873-83.
FR. PIEPER, FA. VAN DE KLUNDEKT,Regulation of vimentin expression in cultured epithelial cells Eur. J. Bioctiem. 210, 509-519 (1992)
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