Efficacy of different methods of isolation on growth of ovine preantral follicles

2^nd International Conference on Animal & Dairy Sciences

September 15-17, 2014 Hyderabad International Convention Centre, India

Sumanta Nandi, Shiv Kumar Tripathi, P S P Gupta and S Mondal

Accepted Abstracts: J Veterinar Sci Technolo

Abstract :

Mammalian ovaries contain an elevated number of preantral follicles that are not easily recruitable for in vitro culture up to the terminal stage of differentiation. Attempts have been made to obtain complete development of mammalian preantral follicles in vitro . In the present study the efficacy of different methods for isolation of preantral follicles from ovine ovaries was examined. Preantral follicles were isolated by mechanical dissection of preantral follicle methods, microdissection, enzymatic digestion (Trypsin digestion 1 and 0.5% for 10 or 5 mins), collagenase digestion (0.5 and 0.25% for 2 and 5 mins). Both small pre-antral follicles (SPFs, 100?250 μ m) and large pre-antral follicles (LPFs, 250?450 μ m) were isolated. The trypsin (1%) digestion for 5 minutes resulted in maximum isolation of total (SPFs: 24.0 ± 3.1 LPFs: 13.2 ± 2.4) and viable preantral (SPFs: 18.0 ± 2.5 LPFs: 10.4 ± 2.0) follicles. The preantral follicles were cultured in vitro for 8 (short term) and 28 days (long term culture), respectively, and examined for their growth, survival and antrum formation rates and growth rates of oocytes in cultured preantral follicles. The in vitro growth, survival and antrum formation rates of preantral follicles and growth rates of oocytes in cultured preantral follicles were significantly (p < 0.05) higher when isolated by trypsin digestion (1% for 5 mins) or mechanical isolation compared to other isolation methods.