Modeling In Vitro Culture of Non-Luteinized Mouse Follicular Granulosa Cells
To investigate the luteinization of mouse follicular granulosa cells,we established an in vitro culture system for non-luteinized granulosa cells using the luteinizing hormone receptor(LHCGR)inhibitor BAY899.Progesterone(P4)levels were measured using ELISA,while quantita-tive PCR was employed to determine the expression levels of follicle-stimulating hormone receptor(FSHR),steroidogenic acute regulatory protein(STAR),cholesterol side-chain cleavage enzyme(CYP11A1),and LHCGR genes.Western Blot was conducted to assess STAR protein levels.From 0 hours to 72 hours,there was a significant increase in P4 levels(P＜0.01);mRNA expression of STAR and CYP11A1 also significantly increased(P＜0.01).In contrast,FSHR mRNA expression significantly decreased(P＜0.01),and LHCGR mRNA expression significantly increased(P＜0.01).BAY899 did not affect cell proliferation but continuously suppressed P4 secretion(P＜0.01)and significantly downregulated both STAR gene and protein expression(P＜0.01).In conclusion,during the in vitro culture of mouse granulosa cells,spontaneous luteinization occurs.BAY899 sig-nificantly inhibits this luteinization process.
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