title: Effect of gonadotrophin on cell and matrix retention and expression of metalloproteinases and their inhibitor in cultured human granulosa cells modelling corpus luteum function.
abstract: Granulosa cells were prepared from follicular aspirates obtained at oocyte collection for in-vitro fertilization (IVF) and maintained in culture. Substantial loss of cells from the culture surface occurred in the absence of gonadotrophin when cells were maintained on a thin layer of extracellular matrix (ECM) using a defined, serum-free medium. This cell loss was clearly and significantly reduced in the presence of human chorionic gonadotrophin (HCG) by days 4-6 of culture, and occurred in conjunction with loss of ECM. Analysis of culture medium by zymography using gelatin as substrate demonstrated the presence of metalloproteinases (MMP), MMP-9 (gelatinase B) appearing as the predominant band. Measurement of overall gelatinase activity in culture media revealed a progressive fall in gelatinase expressed on a per cell basis in media from HCG-treated cultures and this was less marked in controls. This suppression of gelatinase activity was consistent with an observed increase in production of tissue inhibitor of metalloproteinase-1 (TIMP-1) by HCG-treated cells, which was significant by days 6-8 of culture. We speculate that stabilization of the ECM may be an important aspect of HCG action in the corpus luteum.
authors.affiliation: University of Southampton, Princess Anne Hospital, UK.
authors.forename: K E
authors.initials: KE
authors.lastname: Aston
authors.forename: A
authors.initials: A
authors.lastname: Stamouli
authors.forename: E J
authors.initials: EJ
authors.lastname: Thomas
authors.forename: S
authors.initials: S
authors.lastname: Vyas
authors.forename: J P
authors.initials: JP
authors.lastname: Iredale
authors.forename: M J
authors.initials: MJ
authors.lastname: Arthur
authors.forename: M C
authors.initials: MC
authors.lastname: Richardson
mesh_terms: D002478:Cells, Cultured; D006063:Chorionic Gonadotropin / Q000494:pharmacology*; D017364:Collagenases / Q000378:metabolism; D003340:Corpus Luteum Maintenance; D003470:Culture Media; D004247:DNA / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D005109:Extracellular Matrix; D005260:Female; D018093:Gelatinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D006023:Glycoproteins / Q000378:metabolism*; D006107:Granulosa Cells / Q000187:drug effects* / Q000201:enzymology*; D006801:Humans; D020780:Matrix Metalloproteinase 9; D061965:Matrix Metalloproteinase Inhibitors; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D011247:Pregnancy; D018709:Statistics, Nonparametric; D019714:Tissue Inhibitor of Metalloproteinases
chemical_list: D006063:Chorionic Gonadotropin; D003470:Culture Media; D006023:Glycoproteins; D061965:Matrix Metalloproteinase Inhibitors; D019714:Tissue Inhibitor of Metalloproteinases; D004247:DNA; D017364:Collagenases; D018093:Gelatinases; D008666:Metalloendopeptidases; D020780:Matrix Metalloproteinase 9