Influence of protein and hormone supplementation on in vitro maturation and fertilization of domestic cat eggs.
Immature eggs were recovered from freshly excised ovaries from domestic cats, and initially 931 eggs with a compact cumulus and uniform cytoplasm were cultured in 1 of 15 treatments. Eagle's minimum essential medium containing glutamine and pyruvate was supplemented with 5% (v/v) fetal calf serum (FCS), 4 mg bovine serum albumin (BSA) ml-1 or 2 mg polyvinyl alcohol ml-1 (PVA; non-protein control). Within each of these supplement groups, eggs were cultured with: no hormone; LH+FSH; LH+FSH+oestradiol; or LH+FSH+oestradiol+progesterone. After incubation for 52 h, eggs were inseminated with conspecific fresh spermatozoa, cultured and examined for stage of meiotic maturation and in vitro fertilization (IVF). There were fewer (P < 0.05) eggs maturing to metaphase II in vitro in FCS compared with BSA or PVA, the last two treatments producing similar (P < 0.05) results. Gonadotrophins in concert with oestradiol or oestradiol+progesterone improved the incidence of maturation (P < or = 0.01) compared with no added hormones. The incidence of fertilization and cleavage in vitro ranged from 5.2 to 33.9% and varied (P < 0.05) with hormone subtreatment. Adding FSH+LH+oestradiol consistently increased the incidence of IVF approximately twofold compared with controls with no added hormones. Although it inhibited the ability of eggs to reach metaphase II, FCS in the presence of gonadotrophins and oestradiol allowed > 60% of mature eggs to fertilize in vitro (P < 0.05, compared with PVA and BSA). Inhibitory effects on egg maturation were further evaluated by testing four FCS batches from three commercial sources against BSA.(ABSTRACT TRUNCATED AT 250 WORDS)
D000818:Animals; D001798:Blood Proteins; D002415:Cats; D002478:Cells, Cultured; D003470:Culture Media; D004958:Estradiol; D005260:Female; D005307:Fertilization in Vitro / Q000379:methods*; D005640:Follicle Stimulating Hormone; D012739:Gonadal Steroid Hormones; D006065:Gonadotropins, Pituitary; D007986:Luteinizing Hormone; D008297:Male; D009865:Oocytes; D011374:Progesterone
D001798:Blood Proteins; D003470:Culture Media; D012739:Gonadal Steroid Hormones; D006065:Gonadotropins, Pituitary; D011374:Progesterone; D004958:Estradiol; D007986:Luteinizing Hormone; D005640:Follicle Stimulating Hormone