猪冷冻精液体外受精研究

Studies on in vitro Fertilization of Porcine Frozen Semen

为简化猪体外受精过程中精液的来源，避免不同公猪精液和不同采精时间对猪体外受精的影响，本研究采用5mL大管冷冻精液进行体外受精（IVF），研究IVF过程中的各项参数。结果表明：精卵共孵2～8h对多精人卵率影响不大（P〉0．05），2-细胞率随着共孵时间的延长而增加，8h时达到72．8％，显著高于2h组（60．5％，P〈0．05），桑椹胚和囊胚发育率以6h组最高，分别达46．1％和3．48％。多精人卵率随精子浓度的增加而增加，且各组之间差异显著（P〈0．05），2-细胞率也以10^8组（81．2％）最高，且显著高于10^5组（63．7％）（P〈0．05），同时10^7组和10^8组分别获得了8．11％和3．13％的囊胚发育率。以NCSU-23作为成熟培养液，并部分去除卵丘细胞的IVF卵裂率和囊胚率最高，达77．1％和7．33％。冻精和新鲜精液的IVF，无论是在卵裂率，还是桑椹胚或囊胚率上均无显著差异（P〉0．05）。

In order to simplify the source of semen of in vitro fertilization（IVF）, and avoid the effects by different boars and different semen collection time, we used frozen semen by 5 mL- straw to do the IVF. The results showed that there were no significant differences in the polyspermy rates among the 2-8 h oocyte-sperm incubation time（P〈0. 05）. The 2-cell rate was improved along with the increasing of oocyte-sperm incubation time, and 8 h group （72.8%） was much higher than the 2 h group（60.5%, P〈0. 05）. The morula and blastocysts rates of the 6 h group were the highest and got 46.1% and 3.48%, respectively. Polyspermy rate was also increased with the increasing of sperm concentration in the IVF （P〈0.05）. The 2-cell rate of 108 group（81.2%） was the highest, which was much higher than the 105group. The 107 and 106 group got 8.11% and 3.13% blastocyst rate, respectively. Using NCSU-23 as the in vitro maturation（IVM） medium and matured oocytes partly denuded cumulus cells could get the highest cleavage and blastocyst rates（77.1% ,7.33%） among the four groups. The comparation between the frozen semen and fresh semen in the IVF showed that there were no significant differences in the cleavage, morula and blastocyst rates（P〉0. 05）.