Recombinant luteinizing hormone as a survival and differentiation factor increases oocyte maturation in recombinant follicle stimulating hormone-supplemented mouse preantral follicle culture.
The effects of recombinant luteinizing hormone (rLH) and recombinant follicle stimulating hormone (rFSH) both separately and together were analysed on the in-vitro maturation of a well-defined class of mouse early preantral follicles. Metaphase-II (MII) oocytes were only obtained when rLH or rFSH or a combination of both hormones was added to a standard rich culture medium at constant doses throughout the culture period. Without gonadotrophins only 8% of the follicles reached day 12 of culture. Follicle survival was optimal (between 90 and 100%) when rFSH was used; significantly fewer follicles were kept intact when rLH was added as the only supplement (54%; P < 0.001). Theca cells played a fundamental role in follicle survival when rLH was the only supplement. Presence of rLH significantly influenced the formation of antral-like cavities: on day 12, 59% of follicles cultured with rLH + rFSH had a large cavity, as compared with only 21% in cultures with rFSH only (P < 0.0001). The rHCG stimulated germinal vesicle breakdown rate from the different gonadotrophin-supplemented cultures on day 12 was not significantly different (FSH: 89%; LH: 70%; FSH/LH: 83%). However, addition of rLH to rFSH did significantly improve (P < 0.05) the completion of the first meiotic division up to the MII stage (FSH: 46%; LH: 39%; FSH/LH: 76%). Interfollicle differences in steroid production were very large and depended on the presence of theca cells and on the kind of gonadotrophin supplemented. Without theca cells, oestrogen concentrations in rFSH-supplemented cultures were 100 times lower than their theca-containing counterparts during the entire culture period. From culture day 9 onwards most conditioned media had increased basal progesterone concentrations. This study points up the in-vitro role of LH on the differentiation of follicle-like structures and the oocyte's meiotic maturation and suggests an important role for theca cells in providing modulators of in-vitro gonadotrophin action.
Cortvrindt
R
R
Centre for Reproductive Medicine, University Hospital and Medical School, Dutch-speaking Brussels Free University, Belgium.
Hu
Y
Y
Smitz
J
J
D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D006063:Chorionic Gonadotropin / Q000494:pharmacology; D046508:Culture Techniques; D004347:Drug Interactions; D004958:Estradiol / Q000378:metabolism; D005260:Female; D005640:Follicle Stimulating Hormone / Q000008:administration & dosage / Q000494:pharmacology; D007986:Luteinizing Hormone / Q000008:administration & dosage / Q000494:pharmacology*; D008540:Meiosis / Q000187:drug effects; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D009865:Oocytes / Q000166:cytology / Q000187:drug effects* / Q000254:growth & development*; D006080:Ovarian Follicle / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D011374:Progesterone / Q000378:metabolism; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000494:pharmacology
D006063:Chorionic Gonadotropin; D011994:Recombinant Proteins; D011374:Progesterone; D004958:Estradiol; D007986:Luteinizing Hormone; D005640:Follicle Stimulating Hormone