Monolayer culture of human endometrium: Methods of culture and identification of cell types

Abstract

Monolayer cultures can be established from human endometrial tissue after enzymatic dispersal into isolated glands or single cells. Three cell types that have distinct morphology by light and electron microscopy are observed in the resulting primary cultures. One cell type, an elongated spindle cell, is similar in appearance to fibroblasts derived from other tissues. A second cell type forms colonies of tightly cohesive cells, ranging in shape from oval to polygonal. These cells have typical organelles and junctional complexes characteristic of epithelial cells from the endometrium. The third cell type assumes a pavement-like appearance composed of polygonal cells when viewed by phase contrast microscopy, but lacks distinctive ultrastructural features of epithelial cells. These cells in culture resemble the endometrial stromal cell, the predominant cell type of the human endometrium in vivo. The epithelial cell does not survive subculturing but the other two cell types can be passaged through several generations and can be stored in liquid nitrogen and subsequently returned to culture.

Isolation and Culture of Bovine Endometrial Epithelial Cells in a Serum-Free Culture System.

“…I t h a s b ee n w el l de m o n s t r a t ed t h a t t h e production of PGF2α from epithelial cells is increased by stimulation with OT secreted from the corpus luteum and neurohypophysis following luteolysis [2]. However, endometrial stromal cells do not respond to OT [6][7][8]. Endocrinological studies of the bovine uterus have often used uteri in the early estrous stage.…”

“…The initial PGF2α that is released triggers the release of additional OT from the corpus luteum by a positive feedback loop [4,5]. Epithelial cells respond to OT, but stromal cells do not [6][7][8]. Thus, to investigate function of the cultured uterine epithelial cells, the production of PGF 2α in response to OT is useful as a functional marker.…”

Regulation of functional steroid receptors and ligand-induced responses in telomerase-immortalized human endometrial epithelial cells

“…These basal glandular epithelial cells provide a recurrent source for the cellular restitution of the endometrial epithelial lining during the proliferative phase of the cycle. In isolated primary EECs, E2 was unable to sustain telomerase activity, which has been reported to cease within 8 days of culture resulting in the senescence of primary EECs (Varma et al 1982, Tanaka et al 1998.…”

Culture of epithelial and stromal cells of guinea-pig endometrium and the effect of oestradiol-17 on the epithelial cells

“…Effects of oestradiol-17ß on cytosolic progesterone receptor amounts in epithelial cell culture A monolayer of epithelial cells was obtained after 5 days in growth medium; this experiment by phase-contrast microscopy, during cell dissociation, show that the method of successive centri¬ fugation described by Varma et al (1982) is more efficient than that of filtration across a 38 µ stainless-steel grid used by Fleming & Gurpide (1981). The integrity of the glands after digestion by collagenase is better preserved in the first method and the plating efficiency seems to depend on this integrity.…”

“…Cells were dissociated according to the method of Satyaswaroop, Bressler, de la Pena & Gurpide (1979) as modified by Varma et al (1982). The endometrial fragments were incubated in 0-25% (w/v) collagenase, dissolved in CMF-HBSS buffered to pH 7-4 with 20 mM-Hepes, for 2 h at 37°C.…”