Isolation, in vitro maturation, and fertilization of germinal vesicle oocytes obtained from the intact murine ovary

Abstract

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)

Effect of Human Endothelial Progenitor Cell (EPC)- or Mouse Vascular Endothelial Growth Factor-Derived Vessel Formation on the Survival of Vitrified/Warmed Mouse Ovarian Grafts

“…For IVF or intracytoplasmic sperm injection (ICSI), the cumulus-enclosed oocytes were matured in vitro in 50 mL TCM-199 containing 10% fetal bovine serum, 0.0075 IU/mL follicle-stimulating hormone (FSH; Gonal-F; Serono, Modugno Bari, Italy), 0.5 IU/mL hCG (Ovidrel, Serono), and 1 mg/mL E2 (Sigma) in a 5% CO 2 atmosphere for 7 to 9 hours. In our study, for the purpose of obtaining a better oocyte quality, we used E2 and FSH that related to maturation of oocyte cytoplasm for in vitro maturation process as in Randall et al 32 For IVF, capacitated spermatozoa (1-2 Â 10 6 /mL) were incubated with cumulus-enclosed oocytes in Quinn Advantage Fertilization medium (Sage) for 6 hours, and the cumulus cells were removed by repeated pipetting through a glass pipette. The ICSI was performed using a micromanipulator (Narishige, Tokyo, Japan) mounted on an inverted microscope (Nikon Corp) with a Piezo microinjection system (Prime Tech Ltd, Ibaraki-ken, Japan).…”

Null mutation of the prolactin receptor gene produces multiple reproductive defects in the mouse.

“…Generally, a higher level of prolactin is seen in follicles containing mature oocytes capable of being fertilized (Laufer et al 1984;Bohnet et al 1985;Reinthaller et al 1987a,b;Gonen and Casper 1989;Seibel et al 1989;Oda et al 1991;Yoshimura et al 1991), although others find no relationship (Messinis and Templeton 1987) or suggest a negative effect (Irahara et al 1991). Using immature eggs collected from mouse ovaries, prolactin was found to increase the rate of germinal vesicle breakdown and subsequent fertilization and correct development in vitro (Randall et al 1990). The failure of a significant proportion of eggs to undergo germinal vesicle breakdown within maturing PRLR"'^" follicles demonstrates directly the important influence of prolactin and its receptor on oocyte maturation or atresia.…”

“…Divergent effects of PRL on the rate of implantation development of mouse embryos have been reported (Yoshida et al 1987;Yohkaichiya et al 1988;Randall et al 1990). The present studies exclude the absolute requirement for an oocyte PRLR in pre-and postimplantation embryonic development supporting previous investigations Dodds et al 1990) and indicating that the defect must reside in the environment in which the embryo develops.…”

A Preliminary Study of the Effect of Growth Hormone on Mouse Preimplantation Embryo Development in vitro

“…There is evi dence for endocrine regulation of preimplantation em bryonic metabolism in vitro. Prolactin, which resembles GH structurally, increases early embryonic development of in vitro matured germinal vesicle oocytes in inbred mice [3]. Insulin, which binds IGF-I receptors [29], regu lates growth and differentiation in early chick embryos [30], affects glucose metabolism [31] and stimulates gly cine transport [32] and protein synthesis [29] in mouse embryos.…”

“…Ethylenediaminetetraacetic acid (EDTA) has been reported to improve the development of outbred mouse embryos [ 1], as well as the development of mouse embryos derived from oocytes matured in vitro [2], Development of the latter is also positively affected by follicle-stimulating hormone (FSH) or luteinizing hor mone (LH) [2]. Prolactin, which resembles growth hor mone (GH) structurally, promotes early embryonic devel opment of in vitro matured germinal vesicle oocytes in inbred mice [3], Co-culturing of two-cell mouse embryos with a mouse oviductal cell suspension allows embryos to retain cleavage rates and viability comparable to those in vivo [4], Recent evidence about the involvement of GH in ovarian development has caused a reconsideration of its role in reproductive physiology [5] from follicular matura tion to embryonic development.…”