title: Growth and maturation of oocytes in vitro.
abstract: The development of technologies to grow and mature oocytes from the most abundant primordial follicles holds many attractions for clinical practice, animal production technology and research. However, despite much research attention, it has proved difficult to grow follicles from early stages to maturity in vitro, as relatively little is known about the biology of oogenesis. It is clear that throughout oocyte development in vivo, follicle cell support is fundamental to provide the germ cell with nutrients and growth regulators to ensure progression through the protracted growth phase. Conversely, the oocyte actively promotes growth and differentiation of the follicular cells. Both of these characteristics must be mimicked in vitro. Replication of the normal follicular growth span from the primordial to Graafian follicle stages and the changes in the trophic requirements of the cells, cellular interactions, morphogenesis and the sheer increase in bulk as the antrum forms present major challenges for follicle culture technology. These observations could explain why methods that have proved successful for the culture of isolated rodent follicles are unable to support the growth of larger human and ruminant follicles in vitro and are incompatible with the requirements for primordial follicle growth activation. At present, the best option available for the complete growth and maturation of oocytes in vitro is to develop an extended multistage culture strategy which will provide a complex support system that closely resembles the ovary in vivo. In an attempt to achieve this goal primordial follicle growth is first initiated and maintained to the preantral stages through the culture of thin slices of ovarian cortex. The isolation and continued culture of these preantral follicles will support antral cavity formation and the induction of differentiated function in the somatic cell compartment. Finally, after exposure to an appropriate steroid milieu in vitro it should be possible to induce nuclear and cytoplasmic maturation in the fully grown oocytes. The prospects of succeeding at each stage, and of finally producing a fertile gamete, are likely to be increased by preserving cellular interactions and the phenotype of follicle cells as these provide the physiological environment in which oocytes develop. Although the technology for the in vitro maturation (IVM) of fully grown oocytes has been exploited successfully in ruminants, in human assisted reproduction IVM is still experimental as the efficiency of IVM is low and only a small number of pregnancies and live births have been reported. Thus, although complete in vitro growth and maturation may be achieved eventually, immediate goals must include the optimization of methods for isolating and culturing oocytes at both ends of the size spectrum and the full evaluation of the normality of the oocytes grown for extended periods in vitro.
authors.affiliation: Academic Unit of Paediatrics, Obstetrics and Gynaecology, School of Medicine, University of Leeds, Leeds LS2 9NS, UK. H.M.Picton@leeds.ac.uk
authors.forename: H M
authors.initials: HM
authors.lastname: Picton
authors.forename: M A
authors.initials: MA
authors.lastname: Danfour
authors.forename: S E
authors.initials: SE
authors.lastname: Harris
authors.forename: E L
authors.initials: EL
authors.lastname: Chambers
authors.forename: J
authors.initials: J
authors.lastname: Huntriss
mesh_terms: D000818:Animals; D002417:Cattle; D018929:Cell Culture Techniques / Q000379:methods; D002478:Cells, Cultured; D046508:Culture Techniques / Q000379:methods; D004622:Embryo, Mammalian / Q000166:cytology; D005260:Female; D006801:Humans; D008322:Mammals / Q000502:physiology*; D009865:Oocytes / Q000166:cytology / Q000502:physiology*; D009866:Oogenesis; D006080:Ovarian Follicle