Morphogenetic reaggregation and luteinization of mouse preantral follicle cells

Small (60-90 micrometer) and large (100-130 micrometer) preantral follicles were isolated from adult mouse ovaries by a collagenase-dissection technique. These follicles were composed of resting oocytes surrounded either by granulosa cells, only, or by granulosa and undifferentiated theca cells. Further enzymatic dissociation of primary follicles yielded monodisperse cells characterized by abundant rough endoplasmic reticulum, microfilament-rich pseudopodia and only scant lipid droplets. These cells reaggregated, when explanted in stationary culture, forming epithelial cords and structures macroscopically reminiscent of native ovarian follicles. Anticipated association of follicular cells in epithelial-like monolayers was rare (less than or equal to 10% of all cultured cells). Formation and growth of both follicle-like (FLS) and cord-like (CLS) structures occurred within 24 hours of culture, continued for 14 days, and was inhibited by cytochalasin B, but not by neuraminidase. FLS and CLS, as well as cell monolayers, underwent luteinization, as indicated by the presence in the culture medium of radioimmunoassayable progesterone and by frequent cytological features suggestive of active steroidogenesis. The present report indicates that (a) specific cell affinities exist among immature follicular cells which may play a role in folliculogenesis; and (b) follicular cells are endowed, from their early developmental stages with intrinsic steroidogenic capabilities which become phenotypically expressed after escape from the intraovarian environment.