A technique for obtaining basal corneal epithelial cells.
A technique has been developed for obtaining a cell suspension enriched (89%) in basal corneal epithelial cells. Eleven millimeter corneal buttons were removed and placed in culture medium containing low (10 microM) calcium. The posterior half of the stroma was removed with forceps. Three superficial cuts were made with a Bard-Parker blade on the anterior half of the cornea, which was then incubated for 18 hr at 35 degrees C. Nonadherent cells were brushed off after the incubation and basal cells were harvested after a 1-hr incubation in Dispase II. Cell viability estimated by Erythrocin beta exclusion was 90%. Further evidence of viability was that the cells adhered to their native substrate, the denuded basal lamina. The authors protocol provides a method for analyzing the biochemistry of a known population of epithelial cells and makes available a defined source of cells for culture.
D000818:Animals; D001485:Basement Membrane / Q000166:cytology; D002448:Cell Adhesion; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D003315:Cornea / Q000166:cytology*; D003470:Culture Media; D004847:Epithelial Cells; D008855:Microscopy, Electron, Scanning; D011817:Rabbits
D003470:Culture Media