Tesarik J
Centre for Reproductive Biology and Medicine, American Hospital of Paris, Neuilly sur Seine, France.
Hum Reprod. 1993 Aug;8(8):1245-50. doi: 10.1093/oxfordjournals.humrep.a138235.
This study was undertaken to assess the potential of aged human oocytes from failed in-vitro fertilization attempts as a model for the study of fertilization events after subzonal sperm insertion (SUZI). Criteria of aged oocyte suitability for this purpose were (i) the absence of nuclei, (ii) the presence of a polar body, (iii) the absence of cell division or fragmentation, (iv) marked ooplasmic contraction in hyperosmotic medium, and (v) rapid ooplasmic relaxation after returning into normo-osmotic medium following the micromanipulation. Micro-injection techniques were essentially the same as for SUZI with fresh oocytes. Oocytes that fused with the micro-injected spermatozoa developed pronuclei of typical internal structure. However, the number of pronuclei was often higher than that theoretically expected if each sperm nucleus incorporated into the oocyte gave rise to a single pronucleus. Thus, the use of aged oocytes implies the need for a specific method to assess the frequency of fusion in sperm samples examined. The results suggest that the method described here can be applied in a preliminary diagnostic test before a therapeutic SUZI attempt and in studies aimed at the optimization of sperm treatment protocols to increase the fusion capacity of subzonally inserted spermatozoa.
本研究旨在评估来自体外受精失败的老龄人类卵母细胞作为研究透明带下精子注射（SUZI）后受精事件模型的潜力。老龄卵母细胞适用于此目的的标准为：（i）无细胞核；（ii）有极体；（iii）无细胞分裂或破碎；（iv）在高渗培养基中有明显的卵质收缩；（v）显微操作后回到等渗培养基中卵质迅速松弛。显微注射技术与对新鲜卵母细胞进行SUZI时基本相同。与显微注射精子融合的卵母细胞形成了具有典型内部结构的原核。然而，如果每个进入卵母细胞的精核都产生一个原核，原核的数量通常会高于理论预期。因此，使用老龄卵母细胞意味着需要一种特定方法来评估所检测精子样本中的融合频率。结果表明，此处描述的方法可应用于治疗性SUZI尝试前的初步诊断测试以及旨在优化精子处理方案以提高透明带下插入精子融合能力的研究中。
