人类精子空泡及中心体形态对ICSI患者胚胎发育的影响

Impact of the vacuoles and the shape of the sperm midpiece for ICSI on embryos development of patients undergoing ICSI

目的利用精子放大系统将精子放大6000倍后，观察精子是否含空泡及精子中段形态，探讨精子空泡及中心体形态对行卵泡浆内单精子显微注射（ICSI）患者的胚胎发育的影响。方法57例来自成都锦江生殖中心因男方因素行ICSI患者，常规挑选完形态正常的精子（×200-×400）后，再次利用精子放大系统对精子有无空泡和精子中段形态进行观察。根据精子中段形态将精子分为三类，A类：1≤a／b≤1．2；B类：a／b≥1．5；C类：其他不规则形态。有无空泡和精子中段组分为：空泡．A，空泡．B，空泡．C，无空泡．A，无空泡-B，无空泡-C。随后按照常规ICSI注射后培养，并观察胚胎发育情况和患者临床结局。结果空泡．A、B、C3组在2PN率、D3优胚率、可用囊胚率及胚胎冷冻率上均无显著性差异（P〉0．05）；无空泡-A，B，CZ组的可用囊胚率上无显著性差异（P〉0．05），而2PN率（74．48％VS84．62％VS57．14％），卵裂率（96．17％VS97．06％VS80．95％），D3优胚率（35．48％VS39．39％vs9．09％）、D6囊胚形成率（21．05％VS34．78％VS0），胚胎冷冻率（31．25％VS26．92％VS0）均有显著性差异（P〈0．05）。另外A组中有无空泡中，仅仅胚胎冷冻率上具有显著性差异（P〈0．05），B组中2PN率具有显著性差异性（P〈0．05），C组中可用囊胚率和胚胎冷冻率具有显著性差异（P〈0．05），3组中其余指标均无显著性差异（P〉0．0）。结论精子头部无空泡和中心体形态较好的精子注射入卵后，能提高2PN率、D3优胚率、胚胎冷冻率。由于样本量较少，仍然需要较大的样本量来进一步证实，为精子空泡以及中心体对胚胎发育影响的相关研究提供参考。

Objective To clarify if there are any vacuoles at the head of sperm using the motile sperm organelle morphology examination （MSOME） system, and evaluate whether the morphology of the sperm midpiece and the head vacuoles influence further embryo development. Methods Sperms were obtained from 57 patients undergoing ICSI as a result of male infertility in the Center for Reproductive Medicine, Chengdu Jinjiang Hospital for Maternal and Child Health Care. Normal morphology of sperms were selected under a magnification of ×200- × 400 at first, then the selective sperms were observed under high magnification of 6000 ×. According to the shape of sperm medpiece ,there were three classes： （A class： 1 ≤a/b≤ 1.2; B class： a/b≥ 1.5; C class： other）. These sperms were divided into six groups based on the shape of the sperm midpiece and existing vacuoles under 6000 magnification.（vacuoles-A group： 1 ≤a/b≤1.2, there was at least one vacuole exist in the head of sperm vacuoles-B group： a/b≥ 1.5 and there was at least one vacuole exist in the head of sperm; vacuoles-C group： other and there was at least one vacuole exist in the head of sperm; vacuole free-A group; vacuole free-B group; vacuole free-C group） at 6000 magnification. The oocytes of 57 ICSI couples were injected with spermatozoa of six groups. Results There were no significant differences between thevacuoles-A group, vacuolesThere were no statistically significant differences between the vacuoles-A group, vacuoles-B group, vacuoles-C group were observed with regard to rates of fertilization, D3 top quality embryo, blastocyst formation,cryopreservation （P〉0.05）. There were no statistically significant differences in rate of blastocyst formation （/〉〉0.05） between the vacuoles free-A group, vacuole free-B group, vacuole free-C group, the rates of fertilization（74.48 %VS 84.62 %VS 57.14% ）, cleavage （96.17%vs 97.06%vs80.95%）, D3 top quality embryo （35.48% vs 39.39 %vs 9.09%）, D6-blastocyst （21.05%vs34.78%vs0）, cryopreservation（31.25 %vs 26.92% vs 0） were statistically significant different （P〈0.05）. Furthermore, the rate of cryopreservation（14.63 %vs 31.25%）were statistically significant different（P〈0.05）between the vacuoles-A group and vacuoles free-A group. The rate of fertilization（57.14 %vs 84.62%）were statistically significant different （P〈0.05） between the vacuoles-B group and vacuoles free-B group. The rates of blastocyst formation （40.0% vs 0）, cryopreservation （50.00% vs 0） were statistically significant different （P〈0.05） between the vacuoles-C group and vacuoles free-C group. There were no statistically significant difference about other indexes in the three classes sperms. Conclusion A significant improvement in normal fertilization rate and D3 top quality embryo rate and cryopreservation rate was found through selecting sperms based on normal midpiece morphology and no vacuole. This preliminary result need to be further validated by enlarge the sample size