Impact of culture technique on in vitro follicle development

Thesis

Recapitulation of follicle development in vitro is a promising experimental tool that can act both as a model for fundamental research and as a fertility preservation technique. The first live birth was achieved roughly 30-years ago from multi-step neonatal mouse ovary culture that replicated full development from primordial follicle to fertilisation. Nonetheless, translation of this approach to human culture remains limited with only one study demonstrating the development of morphologically normal mature oocytes and poor success rates. Ovarian culture is the gatekeeper of success with this technique, but in vitro follicle development is characterised by reduced theca cell recruitment and growth. Optimisation of ovarian culture is required to enhance follicle development to its full potential. The development of novel culture techniques that can better recapitulate the physiological environment is paramount. In this study, novel culture techniques, roller and RotaTherm, that employ rotary movement are used in attempt to replicate the mechanical forces and fluid shear stress experienced by the ovary within the peritoneal cavity. Development of follicles in mouse ovaries cultured in roller or RotaTherm were compared to stationary membrane culture, the current gold standard for neonatal mouse ovaries, and in vivo age matched controls. Impact of culture technique on follicle health, follicle development, follicle function, extracellular matrix, follicle activation and growth have been analysed through histology and immunochemistry. The results of this study find that rotary culture techniques can support follicle development as well as stationary culture and further improve rates of follicle activation, follicle growth, granulosa layer formation and oocyte growth. However, rotary culture still shows reduced development compared to in vivo controls, highlighting the need for continued optimisation to improve success.

Authors: Boardman, O

• DOI: 10.5287/ora-8nzxxk96e
• Type of award: MSc by Research
• Level of award: Masters
• Awarding institution: University of Oxford
• Language: English
• Keywords: follicle culture; in vitro follicle development; fertility preservation; culture technique; fertility
• Subjects: Reproductive health; Reproductive technology