Serum‐free culture of stromal and functionally polarized epithelial cells of guinea‐pig endometrium: a potential model for the study of epithelial‐stromal paracrine interactions

Abstract

Stromal and glandular epithelial (GE) cells were isolated from guinea-pig endometrium and growth to near confluency (6-8 days) in primary culture on plastic surfaces in a serum-supplemented medium (SSM). The stromal cells were subcultured on plastic dishes and maintained for 72 h in SSM. Then SSM was replaced by a chemically defined medium (CDM) and the stromal cells grown to confluency (5-7 days). The GE cells were subcultured in CDM, on a basement membrane matrix (Matrigel) applied to permeable Millicell-PC filters, and grown to confluency (5 days). Homogeneity of the subcultured endometrial cell populations was ascertained immunocytochemically. The filter-cultured GE monolayers were polarized morphologically, and displayed epithelial-specific specialized structures. These monolayers had functional tight junctions as verified by a measurable transepithelial resistance. The subcultured cell populations were distinguished by an analysis of their cellular and secretory proteins after labelling with [35S]-methionine and analysis by polyacrylamide gel electrophoresis. The filter-cultured GE monolayers allowed identification of the proteins released vectorially in the apical or the basal secretory compartment, thus demonstrating the functional polarization of GE cells in this bicameral culture system. Within the defined conditions of this culture system, the paracrine factors released by the two endometrial cell populations as well as the interplay of stromal-epithelial interactions and ovarian hormones could be investigated.

Establishment of a human endometrial cell culture system and characterization of its polarized hormone responsive epithelial cells

“…Polarized uterine epithelial cells in vitro were investigated in various animal models, such as the rabbit (Mani et al 1991;Winterhager et al 1994), mouse (Jacobs et al 1990(Jacobs et al , 1992, rat Carson et al 1988;Julian et al 1992) and guinea pig (Mahfoudi et al 1991(Mahfoudi et al , 1992(Mahfoudi et al , 1994. All animal studies were performed on epi- thelial cells isolated from uteri of immature animals.…”

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

“…Several studies have reported the serum-free culture of human [14], murine [15], guinea pig [16] and porcine [12] epithelial cells. However, there have been no studies on serum-free culture of bovine endometrial epithelial cells.…”

Effect of progesterone on proteins vectorially secreted by glandular epithelial cells of guinea-pig endometrium: modulation by homologous stroma

“…The recent development of a culture model for guinea-pig endometrial glandular epithelial cells in a bicameral system (Mahfoudi et al, 1992) (Mahfoudi et al, 1991a) and this steroid stimulated inorganic sulfate incorporation into the cells (Mahfoudi et al, 1991b (Bell et al, 1987). This progesterone-dependent protein, called PP14 (Rutanen et al, 1987), or o2-PEG (Bell et al, 1985) or progestagen-dependent endometrial protein (Joshi et al, 1980), related to ß-lactoglobulins, could function as a ligand-carrier (Bell et al, 1987).…”

“…The endometrium is an epithelium/mesenchyme stromal structure that has an active role in producing autocrine and paracrine factors that modulate the physiology of steroid-dependent tissue (Inaba et al, 1988;Brigstock et al, 1989; Tabibzadeh, 1991). It has been suggested that at the time of blastocyst attachment the uterine epithelium may influence stromal decidualization via soluble factors (Lejeune et al, 1981;Tamada et al, 1990Tamada et al, , 1991 (Inaba et al, 1988;Hohn et al, 1989) (Glasser et al, 1988) and rabbits (Mani et al, 1991) (Chaminadas et al, 1986) and cultured and subcultured as decribed by Mahfoudi et al (1992) Mahfoudi et al (1991a).…”

“…3, lane C), the electrophoretic profiles of apical and basal secretions showed bands that represented markers of each compartment as described by Mahfoudi et al (1992 (Fig. 3).…”