Isolation culture and inmunohistochemical identification on vaginal epithelial cells

Abstract

Vaginal epithelial cells from 250-g Wistar rats were cultivated in plastic dishes without feeder layer support of cells of mesodermic origin. Isolated epithelial cells were obtained from everted vaginas by exposing the epithelium to a solution of 0.5% trypsin and cultivated in a modified Ham's F 12 medium supplemented with fetal calf serum, insulin. hydrocortisone and cholera toxin. In this way. epithelial cells gave rise to epithelioid colonies after 48hr of culture. Infrequent fibroblast contamination was observed. Positive confirmation o í the epithelial origin of the cell colonies was carried out with antikeratin antibodies and immunohistochemical techniques. This cell culture was shown to proliferate and differentiate in vitro. Proliferation has been assayed by the capacity of the cells to incorporate [3H]thymidine and differentiation by the presence in the cell culture of cells possessing disulfide-bonded keratins and a chemically resistant protein envelope. Addition of 17β-estradiol to serum-deprived cell cultures induced an increase in the total protein per dish over the serumless cell cultures, but not reaching the values of serum-treated cultures. This tissue culture system seems to be a suitable model to study effects of hormones, growth factors etc. on proliferation and differentiation of isolated cells.