Proliferative potential and polymorphism of human endometrial stromal cells

The proliferative potential of stromal cells from human endometrium, separated from glands by dispersion in the presence of collagenase and subsequent passage through a sieve, was evaluated by determining the total number of cell doublings achieved when cells were repeatedly subcultured in a 1:2 split ratio in Ham's F-10 medium containing 10% fetal bovine serum (FBS). Fifty doublings or more (up to 100) were observed in 8 of the 26 specimens (30%) which we examined. This number of doublings is high for cells obtained from adult tissues and may in part reflect the unusually great proliferative capacity of human endometrium when compared to that of other tissues. The shape of the stromal cells depended on the medium in which they were originally cultured. Cells cultured in Ham's F-10 medium containing 10% FBS showed the typical fibroblastic morphology at confluence; they appeared elongated or spindle-shaped and formed monolayers. In contrast, cultures in CMRL-1066 medium in the presence of 10% FBS appeared polygonal or stellate-shaped and also formed monolayers. In about 50% of the cultures in CMRL-1066 medium we observed fibroblast-shaped cells that superficially resembled cells grown in Ham's F-10 medium, but were able to form dome structures. In some cultures the regions of prominent overgrowth were macroscopically visible. Switching media during later passages did not reverse the shape of the cells obtained in CMRL-1066 medium or that of the fibroblast-shaped cells in Ham's F-10 medium, suggesting either that the growth of a subpopulation was favoured early during cellular adaptation to primary culture or that there was a single cell population whose phenotype was determined early in culture and then no longer responded to medium factors. Examination of the cytoskeleton after visualization with rhodamine-labelled phalloidin revealed that the arrangement of the microfilaments corresponded to the cell shape observed in living cells under the phase contrast microscope. Distinct changes in morphology were observed when primary stromal cell cultures in CMRL-1066 medium were exposed to progesterone, indicating that progestins may affect cytoskeletal proteins.