Isolation, in vitro maturation, and fertilization of germinal vesicle oocytes obtained from the intact murine ovary.
The purpose of this investigation was to attempt to develop a process, utilizing a murine model, which would allow more efficient harvesting from the intact ovary and maturation in vitro of germinal vesicle (GV) oocytes. The recovery process yielded 25.5 +/- 4.5 (mean +/- SE) cumulus-free GV oocytes per animal. Treatment groups included culture medium (CM) supplemented with either estradiol (E2), follicle stimulating hormone (FSH), human chorionic gonadotropin (hCG), or prolactin (PRL). Among the hormone-free controls 83.2 +/- 1.6% of oocytes underwent GV breakdown, whereas 25.3 +/- 2.6% developed to the first polar body stage (PB-1) following 18 hr of incubation (n = 29 trials). Oocytes progressing to the PB-1 stage were inseminated in vitro. In vitro fertilization (IVF) of pooled in vitro matured (IVM) PB-1 oocytes (judged by two-cell formation) was 19.9%, which was significantly lower than in the group of in vivo matured oocytes (74.4%). E2 significantly increased the percentage of GV breakdown (control, 76.8 +/- 2.5%; E2 at 10 ng/ml, 92.9 +/- 2.5%, P less than 0.001; E2 at 100 ng/ml, 93.7 +/- 2.1%, P less than 0.001; and E2 at 1 micrograms/ml, 86.7 +/- 3.3%, P less than 0.05) but not PB-1 formation. Neither FSH nor hCG significantly increased GV breakdown or PB-1 formation.(ABSTRACT TRUNCATED AT 250 WORDS)
Randall
G W
GW
Indian Path Medical Center, Fertility Resources Center, Kingsport, Tennessee 37660.
Awadalla
S G
SG
Shivers
C A
CA
D000818:Animals; D002469:Cell Separation; D002478:Cells, Cultured; D006063:Chorionic Gonadotropin / Q000032:analysis / Q000494:pharmacology; D003470:Culture Media / Q000032:analysis / Q000494:pharmacology; D004958:Estradiol / Q000032:analysis / Q000494:pharmacology; D005260:Female; D005306:Fertilization / Q000187:drug effects / Q000502:physiology*; D005307:Fertilization in Vitro / Q000379:methods; D005640:Follicle Stimulating Hormone / Q000032:analysis / Q000494:pharmacology; D051379:Mice; D009865:Oocytes / Q000166:cytology* / Q000187:drug effects / Q000502:physiology; D009866:Oogenesis / Q000187:drug effects / Q000502:physiology*; D010053:Ovary / Q000166:cytology* / Q000187:drug effects / Q000502:physiology; D011388:Prolactin / Q000032:analysis / Q000494:pharmacology
D006063:Chorionic Gonadotropin; D003470:Culture Media; D004958:Estradiol; D011388:Prolactin; D005640:Follicle Stimulating Hormone