title: Secondary follicle growth and oocyte maturation during encapsulated three-dimensional culture in rhesus monkeys: effects of gonadotrophins, oxygen and fetuin.
abstract: BACKGROUND
  An alginate-based matrix supports the three-dimensional (3D) architecture of non-human primate follicles and, in the presence of FSH, permits the in vitro development of pre-antral follicles to the small antral stage, including the production of ovarian steroids and paracrine factors. The current study investigated the ability of gonadotrophins, fetuin and oxygen (O₂) to improve primate follicle growth and oocyte maturation in vitro.
  
  
  METHODS
  Macaque secondary follicles were isolated from the early follicular phase ovaries, encapsulated in a sodium alginate matrix and cultured individually for 40 days in supplemented medium. The effects of recombinant human (rh) FSH (15, 3 and 0.3 ng/ml for high, medium and low FSH, respectively), bovine fetuin (1 or 0 mg/ml) and O₂ (5 or 20% v/v) were examined. Half of the follicles in each culture condition received rhLH on Day 30-40. Follicles that reached antral stage were treated with rh chorionic gonadotrophin for 34 h to initiate oocyte meiotic maturation. Media were analyzed for ovarian steroids and anti-müllerian hormone (AMH).
  
  
  RESULTS
  Improved culture conditions supported non-human primate, secondary follicle growth to the antral stage and, for the first time, promoted oocyte maturation to the MII stage. In the presence of fetuin at 5% O₂, follicles had the highest survival rate if cultured with high or medium FSH, whereas follicles grew to larger diameters at Week 5 in low FSH. Oocyte health and maturation were promoted under 5% O₂. High FSH stimulated steroid production by growing follicles, and steroidogenesis by follicles cultured with low FSH was promoted by LH. AMH biosynthesis was elevated with high compared with low FSH and for longer under 5% O₂ than under 20% O₂.
  
  
  CONCLUSIONS
  This encapsulated 3D culture model permits further studies on the endocrine and local factors that influence primate follicle growth and oocyte maturation, with relevance to enhancing fertility preservation options in women.
authors.affiliation: Division of Reproductive Sciences, Oregon National Primate Research Center, Oregon Health & Science University, Beaverton, OR 97006, USA.
authors.forename: J
authors.initials: J
authors.lastname: Xu
authors.forename: M S
authors.initials: MS
authors.lastname: Lawson
authors.forename: R R
authors.initials: RR
authors.lastname: Yeoman
authors.forename: K Y
authors.initials: KY
authors.lastname: Pau
authors.forename: S L
authors.initials: SL
authors.lastname: Barrett
authors.forename: M B
authors.initials: MB
authors.lastname: Zelinski
authors.forename: R L
authors.initials: RL
authors.lastname: Stouffer
mesh_terms: D000818:Animals; D054304:Anti-Mullerian Hormone / Q000378:metabolism; D018929:Cell Culture Techniques; D005260:Female; D005640:Follicle Stimulating Hormone / Q000494:pharmacology*; D012739:Gonadal Steroid Hormones / Q000378:metabolism; D006062:Gonadotropins / Q000494:pharmacology*; D008253:Macaca mulatta / Q000502:physiology*; D009865:Oocytes / Q000187:drug effects / Q000254:growth & development* / Q000378:metabolism; D006080:Ovarian Follicle / Q000187:drug effects / Q000254:growth & development* / Q000378:metabolism; D010100:Oxygen / Q000494:pharmacology*; D000509:alpha-Fetoproteins / Q000494:pharmacology*
chemical_list: D012739:Gonadal Steroid Hormones; D006062:Gonadotropins; D000509:alpha-Fetoproteins; D054304:Anti-Mullerian Hormone; D005640:Follicle Stimulating Hormone; D010100:Oxygen