本研究旨在建立一套适合水牛腔前卵泡体外生长发育的培养体系。取用来自本地屠宰场的中国沼泽型水牛卵巢,采用梳刮法回收腔前卵泡,以McCoy’s 5a作为基础培养液,分别用微孔板培养法、二维培养法、三维培养法进行体外培养。结果表明:不同培养方法对水牛腔前卵泡的体外发育能力有显著差异。培养至10d,三维培养法的卵泡存活率显著高于微孔板培养法和二维培养法的卵泡存活率(65.05% vs 33.08%,49.52%,P〈0.05);二维培养法的卵泡成腔率为1.91%(2/105),三维培养法的卵泡成腔率为1.94%(2/103),而微孔培养法的卵泡未发现成腔;三维培养法的卵泡直径平均增长显著高于微孔板培养法和二维培养法的卵泡直径增长(13.03±5.37μm vs 7.53±2.26μm,10.27±4.24μm,P〈0.05)。由此可见,三维培养法是水牛腔前卯泡的有效体外培养方法。
A suitable culture system was established for the growth of buffalo preantral follicles in vitro. Buffalo ovaries were obtained from the local slaughterhouse. The comb-scrape method harvested preantral follicles. The base culture medium was McCoy's 5a medium. Preantral follicles were cultured with the micro-plate culture method, two-dimension cultural method and three-dimension cultural method in vitro culture. Effects of culture method on the in vitro development of buffalo preantral follicles were examined. The results showed that different cultural methods resulted in significantly different in vitro development of buffalo preantral follicles. The survival rate of follicles cultured in three-dimension culture method for 10 d was significantly higher than that of in micro-plate and two-dimension culture methods (65.05% vs 33.08%, 49.52% , P 〈 0.05 ). Buffalo antral follicle rates from preantral follicles cultured by three-dimension and two-dimension for 10 d were 1.91% (2/105) and 1.94% (2/103), respectively. But the micro-plate had no antral follicles. The increase in follicle diameter in the three-dimension culture method was also significantly higher than that of micro-plate and two-dimension culture methods ( 13.03 ± 5.37μm vs 7.53 ± 2.26 μm, 10.27 ± 4.24 μm, P 〈 0. 05 ). These results indicate that the threedimension culture method is more efficient for in vitro culture of buffalo preantral follicles.