In Vitro Development of Marmoset Monkey Oocytes by Pre-antral Follicle Culture

✍ Scribed by PL Nayudu; J Wu; HW Michelmann

• Publisher: John Wiley and Sons
• Year: 2003
• Tongue: English
• Weight: 206 KB
• Volume: 38
• Category: Article
• ISSN: 0936-6768
• DOI: 10.1046/j.1439-0531.2003.00398.x

✦ Synopsis

Contents

A technique for in vitro maturation of oocytes from small ovarian follicles of marmoset monkeys (Callithrix jacchus) has been developed. We employed a two‐step culture system for primary follicles (45–85 __μ__m) and a one‐step culture technique for secondary follicles (>85 __μ__m). The two‐step technique started with the culture of stromal tissue fragments for 2 days. Thereafter, mechanically isolated follicles were transferred to a culture system where they attached to the culture surface and grew for up to a further 12 days. Significant growth of the small follicles and their oocytes was only achieved with gonadotrophins in the medium. Oocytes with a mean diameter of 39 __μ__m from follicles <85 __μ__m reached a mean diameter of 90 __μ__m by the end of the two‐step culture. After in vitro maturation, 19% of oocytes from these follicles had progressed to the germinal vesicle breakdown (GVBD). Follicles between 85 and 170 __μ__m in diameter were isolated from the stroma and placed directly in the culture. Oocytes from these follicles had a mean diameter of 64 __μ__m. The maximum size the oocytes reached in culture was related to the age of the females (pre‐pubertal females: 102 ± 1.3 __μ__m; adults: 96 ± 1.4 __μ__m). Twenty‐seven per cent of oocytes from pre‐pubertal ovaries achieved GVBD and nearly two‐thirds of these progressed to polar body stage. From adult ovaries, only 12% progressed to GVBD and one‐third of these to polar body stage. It is possible to develop mature oocytes in vitro from marmoset secondary pre‐antral follicles (>85 __μ__m). From primary follicles, although near full size oocytes were developed, maturation capacity was incomplete.