Stock C E, Bates R, Lindsay K S, Edmonds D K, Fraser L R
Chelsea Hospital for Women, London, UK.
J Reprod Fertil. 1989 Jul;86(2):723-30. doi: 10.1530/jrf.0.0860723.
Oocyte-cumulus complexes were obtained, after induced ovulation, from infertile patients participating in an in-vitro fertilization programme. About 6 h after retrieval and depending on the expansion of the cumulus, 100,000 motile spermatozoa, prepared by a migration-centrifugation method, were added. After 14-18 h incubation at 37 degrees C, oocytes were examined for signs of fertilization (pronuclei and polar body formation) and then removed; spermatozoa remaining in the incubation medium were fixed for transmission electron microscopy. To provide an adequate number of cells for observation, spermatozoa from a minimum of 3-5 oocytes from the same patient were pooled. When sufficient spermatozoa were available after insemination, the remainder of the suspension was incubated at 37 degrees C and fixed along with the corresponding oocyte-incubated sample. In all, 32 sperm samples were assessed and fertilized oocytes were obtained with 29 of these. In the 24 samples in which greater than 100 spermatozoa (mean of 192) could be assessed, 32% of spermatozoa had initiated or completed the acrosome reaction. In the 15 of these 24 samples for which oocyte-free controls were available, 31% of cells were reacting or reacted, compared with 15% of cells (P less than 0.001) in the controls. In the remaining 8 samples, incubated with oocyte-cumulus complexes, less than 100 but greater than or equal to 20 spermatozoa (mean of 42) were assessed and again 32% of spermatozoa were reacted.(ABSTRACT TRUNCATED AT 250 WORDS)
在诱导排卵后,从参与体外受精项目的不孕患者中获取卵母细胞 - 卵丘复合体。取回后约6小时,根据卵丘的扩展情况,添加通过迁移 - 离心法制备的100,000个活动精子。在37℃孵育14 - 18小时后,检查卵母细胞的受精迹象(原核和极体形成),然后取出;将留在孵育培养基中的精子固定用于透射电子显微镜检查。为了提供足够数量的细胞进行观察,将来自同一名患者至少3 - 5个卵母细胞的精子汇集在一起。授精后当有足够的精子时,将悬浮液的剩余部分在37℃孵育,并与相应的卵母细胞孵育样品一起固定。总共评估了32个精子样本,其中29个获得了受精卵母细胞。在24个可评估超过100个精子(平均192个)的样本中,32%的精子已启动或完成顶体反应。在这24个样本中有15个有未与卵母细胞共孵育的对照,其中31%的细胞正在反应或已经反应,而对照中的细胞为15%(P小于0.001)。在其余8个与卵母细胞 - 卵丘复合体共孵育的样本中,评估的精子少于100个但大于或等于20个(平均42个),同样有32%的精子发生了反应。(摘要截断于250字)