Primary Culture and Characterization of Human Ovarian Surface Epithelial Cells

KANOH,Hideo, MATSUI,Yoshiaki, MAKINO,Toshiko, OKUDAIRA,Yoshio

Primary culture and characterization of the human ovarian surface epithelial cells were carried out to make a preliminary trial of the histogenetic consideration of common epithelial tumors of the ovary utilizing in vitro systems. Ovarian surface epithelial cells were harvested from strips of surgically excised ovary using an explant culture method and submitted to studies. The cells appeared flat epithelioid in shape and arranged in a cobblestone pattern. Immunocytochemical stainings by the PAP method revealed that these epithelioid cells were keratin and vimentin positive. The cells grew in the medium of RPMI 1640 with 10% fetal calf serum and the growth rate was accelerated significantly when supplemented with hEGF (10ng/ml) and hydrocortisone (0.5/μg/ml). When epithelioid cells were cultivated on Matrigel coated glass, some morphological changes were brought about by treating them with TPA (10ng/ml) or sodium butyrate (1.5mM). When the cells were cultivated on type I collagen gel, the arrangement of the cells appeared to be mostly monolayer. Grown in Matrigel overlaid on type I collagen gel, the cells grouped and arranged seemingly in spreading branches. Although no malignant transformation has been observed, some morphological changes reminiscent of metaplastic change were seen in some of the cultured cells when the media were treated repeatedly with 5-azacytidine (3μM), the DNA hypomethylating agent. The results reported may help to dome extent to clarify the biological nature of ovarian surface epithelial cells including ovarian tumorigenesis which still remains unknown.

Primary Culture and Characterization of Human Ovarian Surface Epithelial Cells

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