Evidence for the Oligoclonal Origin of the Granulosa Cell Population of the Mature Human Follicle 1

Abstract

The clonality of the granulosa cell population residing in individual mature human ovarian follicles was examined by determining the pattern of X chromosome inactivation. Granulosa cells from 72 follicles were obtained from 9 patients undergoing oocyte harvest for in vitro fertilization. The granulosa cell DNA obtained from each follicle was subjected to the PCR, to amplify a highly polymorphic region of the X-linked human androgen receptor gene, after digestion by the methylation-sensitive HpaII restriction endonuclease, thereby achieving exclusive amplification of the inactive allele. Seventeen of 65 informative follicles (26 +/- 5%) were comprised of granulosa cells exhibiting inactivation of the same X chromosome. At least 1 such follicle was found in 8 of the 9 women sampled. There are 2 possible explanations for these findings: 1) approximately one fourth of all follicles contain a truly monoclonal granulosa cell population; 2) the granulosa cells of a given follicle are derived from a small number of stem cells (3 cells), such that the probability is 0.25 that all 3 stem cells producing the granulosa cell complement of a given follicle have the same X chromosome inactivated by chance. We favor the latter explanation and conclude that the granulosa cell cohort of mature human follicles is oligoclonal.