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.
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.
Mahfoudi
A
A
Unité de Biochimie Hormonale et des Régulations, INSERM U 198, Besançon, France.
Fauconnet
S
S
Bride
J
J
Beck
L
L
Remy-Martin
J P
JP
Nicollier
M
M
Adessi
G L
GL
D000818:Animals; D001485:Basement Membrane; D016764:Cell Polarity; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free; D046508:Culture Techniques / Q000295:instrumentation / Q000379:methods*; D004717:Endometrium / Q000166:cytology* / Q000378:metabolism; D004847:Epithelial Cells; D005260:Female; D006133:Growth Substances / Q000302:isolation & purification / Q000378:metabolism*; D006168:Guinea Pigs
D016895:Culture Media, Serum-Free; D006133:Growth Substances