What we have learned from isolated cells from human ovary?

Abstract

In the ovary, morphodynamics of follicles with cyclic maturation, ovulation and repair occur under the control of various tropic factors. The ovarian functions have been mostly studied by using subhuman primates and non-primate animals because of the limited availability of closely staged human specimens. We have recently established the in vitro culture systems of ovarian surface epithelium (OSE) and granulosa cells of humans, and subsequently developed the immortalization of each cell. The immortalized cell lines may supply us advanced studies on ovarian disorders as well as its physiological functions. On the embryologically putative müllerian potential of coelomic epithelium, endometriosis can be explained by coelomic metaplasia from the peritoneal mesothelium, including OSE. We can microscopically observe a continuity from flat epithelial cells on the ovarian surface or within the cortical inclusion cysts to endometriotic gland cells. The primary human OSE cells exhibited a glandular–stromal structure similar to endometriosis when they were co-cultured with endometrial stromal cells in an estrogen-rich environment. Primary and immortalized OSE cells converted the estrone to estradiol, and expressed the genes for steroidogenic factor-1 (SF-1), p450arom and 17β-HSDs. This character of OSE was, in part, similar to the granulosa cells. One of the immortalized OSE clone produces disseminated tumors mimicking undifferentiated carcinomas in nude mice. Ovarian granulosa cells play a key role in the functional maturation of the entire follicle. The molecular pathways in granulosa cells responsible for the growth, differentiation, and nursing the oocyte are still largely unknown. Our immortalized human granulosa cell line, GC1a, obtained from developing follicles, showed no steroid hormone biosynthesis, and no detectable expression of the genes for StAR or cytochrome p450 enzymes due to the lack of SF-1. Transfected SF-1 elicited estradiol secretion in GC1a cells with concomitant expression of the genes encoding the proteins for gonadal steroidogenesis. The enzymatic activity of 17β-HSD was also achieved by SF-1 transgene. These results indicate that SF-1 controls the gene expression required for steroidogenesis in the human developing follicle. Clinically, immortalized GC1a cells from human origin, with steroidogenic capacity, may serve as a feeder layer for in vitro oocyte maturation. Further investigations of our immortalized OSE and granulosa cells of humans will allow us to clarify whether they have a single progenitor cell.