In vitro culture and in vitro maturation of mouse preantral follicles with recombinant gonadotropins.
OBJECTIVE
To develop an effective method for in vitro maturation of preantral follicles isolated from mice ovarian tissue.


METHODS
Isolated preantral follicles were randomly allocated to designed experimental groups for study.


METHODS
University-based research lab.


METHODS
Healthy, normal mice.


METHODS
Superovulation with pregnant mare serum gonadotropin and hCG.


METHODS
Morphological changes and E(2) production were assessed.


RESULTS
To obtain competent oocytes, preantral follicles must be cultured with medium containing insulin and recombinant gonadotropins (i.e., recombinant FSH and recombinant LH), with a change of medium daily. A high initial recombinant LH or recombinant FSH facilitates E(2) secretion, enhances granulosa cell outgrowth, and has earlier antral formation. However, prolonged culture in high-recombinant LH or recombinant FSH triggers early differentiation and luteinization of granulosa cells, which results in low metaphase II oocyte and blastocyst formation.


CONCLUSIONS
We have developed a culture system that allows the successful maturation of preantral follicles in vitro. The matured follicles are a physiologically functional unit that not only secrete E(2) but also generate competent oocytes. In a special condition, 90% of the cultured follicles survived, 53.5% of them produced MII oocytes, and 50% of the derived MII oocytes were fertilized and reached the blastocyst stage after culture in vitro.
Liu
Hung Ching
HC
The Center for Reproductive Medicine and Infertility, Weill Medical College of Cornell University, 515 71st Street, New York, NY 10021, USA. hcliu@mail.med.cornell.edu
He
Zhiying
Z
Rosenwaks
Zev
Z
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D004958:Estradiol / Q000096:biosynthesis; D005260:Female; D005307:Fertilization in Vitro; D005640:Follicle Stimulating Hormone / Q000494:pharmacology*; D006736:Horses; D007328:Insulin / Q000494:pharmacology; D007986:Luteinizing Hormone / Q000494:pharmacology*; D008297:Male; D051379:Mice; D006080:Ovarian Follicle / Q000166:cytology / Q000187:drug effects / Q000254:growth & development* / Q000502:physiology; D011247:Pregnancy; D011897:Random Allocation; D011994:Recombinant Proteins / Q000494:pharmacology
D007328:Insulin; D011994:Recombinant Proteins; D004958:Estradiol; D007986:Luteinizing Hormone; D005640:Follicle Stimulating Hormone