This study aimed to investigate freezing effects of ovarian tissues on survival of preantral follicles and observing in vitro growing of preantral follicles retrieved from cryopreserved ovarian cortical tissues of a cheetah post-mortem. After 29-hour cold storage, ovarian cortices were cut into small pieces (2.0 x 2.0 x 1.0 mm3) and allocated to be frozen using a passive cooling container (n = 3 pieces) or vitrification (n=3 pieces). After one year of storage, 43 (10/23) and 21% (12/58) follicles isolated from ovarian tissues cryopreserved using a passive cooling device (slow freezing rate) and vitrification, respectively, were viable (positively stained with neutral red).Thereafter, the viable follicles were in vitro grown in a culture medium containing M199 supplemented with growth hormone (GH), follicular-stimulating hormone (FSH), insulin-like growth factor I (IGF-I)and activin A for 7 days. Diameters and diameter gains were examined on Days 0, 3 and 7. Follicle viability was assessed on Days 0 and 7. Diameters of follicles frozen by the slow freezing decreased gradually from 53.5±14.2 μm on Day 0 to 50.9±17.1μm with 2 out of 10 viables, whereas those frozen using vitrification maintained their diameters between 50.7±15.6 μm and 50.5±17.9 μm on Days 0 and 7, respectively, with 2 of 12 viable. In conclusion, the passive cooling container is suggested to perform a slow freezing rate for ovarian tissue cryopreservation. Although the cheetah ovarian follicles obtained from cryopreserved tissues can be grown in vitro for 7 days, optimization of culture medium is required to improve the viability and growing rate.
Faculty of Veterinary Science, Chulalongkorn University
Wongbandue, Grisnarong; Tanpradit, Nae; Thongthainun, Daraka; Thuwanut, Paweena; and Chatdarong, Kaywalee
"Viability and Growth of Preantral Follicles Derived from Cryopreserved Ovarian Tissues of a Cheetah (Acinonyx jubatus) Post-mortem,"
The Thai Journal of Veterinary Medicine: Vol. 43:
3, Article 14.
Available at: https://digital.car.chula.ac.th/tjvm/vol43/iss3/14