Survival, growth, and maturation of secondary follicles from prepubertal, young, and older adult rhesus monkeys during encapsulated three-dimensional culture: effects of gonadotropins and insulin.
A three-dimensional culture system supports the development of primate preantral follicles to the antral stage with appreciable steroid production. This study assessed i) whether in vitro developmental competence of follicles is age dependent, ii) the role of gonadotropins and insulin in supporting folliculogenesis, and iii) anti-Müllerian hormone (AMH) and vascular endothelial growth factor (VEGF) production by growing follicles. Ovaries were obtained from prepubertal, young, and older adult rhesus macaques. Secondary follicles were encapsulated into alginate beads and cultured individually for 40 days in media containing 0.05 or 5  μg/ml insulin, with or without recombinant human (rh) FSH (500  mIU/ml). No follicles survived in the culture without rhFSH. In the presence of rhFSH, survival was lower for follicles from older animals, whereas growth, i.e. follicle diameter, was less by day 40 for follicles from prepubertal animals. The surviving follicles were categorized as no-grow (NG; ≤ 250 μm), slow-grow (SG; 250-500 μm), and fast-grow (FG; ≥ 500  μm) according to their diameters. SG follicles cultured with 5 μg/ml insulin produced more ovarian steroids than those cultured with 0.05  μg/ml insulin by week 5. SG and FG follicles produced more AMH and VEGF than the NG, and levels peaked at weeks 2 and 5 respectively. After 100  ng/ml rh chorionic gonadotropin treatment for 34 h, more healthy oocytes were retrieved from young adults whose follicles were cultured with 5  μg/ml insulin. This culture system offers an opportunity to characterize the endocrine and paracrine function of primate follicles that influence follicle growth and oocyte maturation.
Xu
Jing
J
Division of Reproductive Sciences, Oregon National Primate Research Center, Oregon Health & Science University, 505 Northwest 185th Avenue, Beaverton, Oregon 97006, USA.
Bernuci
Marcelo P
MP
Lawson
Maralee S
MS
Yeoman
Richard R
RR
Fisher
Thomas E
TE
Zelinski
Mary B
MB
Stouffer
Richard L
RL
D000464:Alginates / Q000494:pharmacology; D000818:Animals; D054304:Anti-Mullerian Hormone / Q000032:analysis / Q000378:metabolism*; D004958:Estradiol / Q000032:analysis / Q000378:metabolism; D005260:Female; D005640:Follicle Stimulating Hormone / Q000502:physiology*; D020723:Glucuronic Acid / Q000494:pharmacology; D006603:Hexuronic Acids / Q000494:pharmacology; D066298:In Vitro Techniques; D007328:Insulin / Q000502:physiology*; D008253:Macaca mulatta / Q000502:physiology*; D009865:Oocytes / Q000502:physiology; D006080:Ovarian Follicle / Q000166:cytology / Q000502:physiology*; D011374:Progesterone / Q000032:analysis / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000032:analysis / Q000378:metabolism*
D000464:Alginates; D006603:Hexuronic Acids; D007328:Insulin; D042461:Vascular Endothelial Growth Factor A; D011374:Progesterone; D004958:Estradiol; D054304:Anti-Mullerian Hormone; D020723:Glucuronic Acid; D005640:Follicle Stimulating Hormone