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The basic study of the electrofertilization using an electrofusion technique

电融合技术电受精的基础研究

基本信息

批准号：
05671346
负责人：
SENGOKU Kazuo
金额：
$ 1.02万
依托单位：
ASAHIKAWA MEDICAL COLLEGE
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1993
资助国家：
日本
起止时间：
1993 至 1994
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05671346/
关键词：
electrofusion subzonal insemination

项目摘要

We studied the probability of the successful fertilization using an electrofusion methods, and obtained the following results.1.In the experiments that three to ten mouse sperms were injected into the perivitelline space of each mouse oocytes, the best results of sperms attachment, fertilization were obtained after application of dc of 3.5KV/cm, 10musec followed by ac pulses (2MHz, 100V/cm, 20sec each). After pulse application, 25.5% of the oocytes were fertilized. However, 90% of the oocytes showed the polyspermic fertilization and 15.5% of the oocytes were degenerated.2.Oocyte activation prior to application of fusion pulse is known to facilitate oocyte-sperm fusion. When the oocytes were electroactivated by exposures to ac pulse (2MHz, 50V/cm, 10sec) and followed by dc (1.5KV/cm, 50musec), about 70% of the oocytes were activated. Twenty percents of the oocytes stimulated with electoactivation developed to the pronucleus stage after the application of the electrofusion. However, all these oocytes were arrested at the PN stage.3.When a single mouse sperm was injected into the perivitelline space and applied an electrofusion technique, 12.5% of these oocytes were fertilized and developed to the PN stage.4.In the trial using human sperms and hamster oocytes, 3.0% of the hamster oocytes were ferilized under the same conditions of electrofusion. However, all oocytes were arrested at the PN stage and 15.0% of the oocytes were degenerated.Although the success rate is very low and more study about the best conditions for electrofusion is needed, our preliminary results may offer important prospects of the treatment of the patients with sperm dysfunction.

1.将3 ~ 10个小鼠精子分别注入卵母细胞的卵周隙中，以3.5KV/cm，10 μ sec的直流电和2 MHz，100 V/cm，20 sec的交流脉冲，精子附着、受精的效果最好。脉冲处理后，25.5%的卵母细胞受精。然而，90%的卵母细胞显示多精受精，15.5%的卵母细胞变性。2.在应用融合脉冲之前激活卵母细胞有助于卵母细胞-精子融合。用交流电（2 MHz，50 V/cm，10 sec）和直流电（1.5KV/cm，50 musec）激活卵母细胞，约70%的卵母细胞被激活。20个经电激活的卵母细胞在电融合后发育到原核期。将小鼠精子注入卵周间隙，采用电融合技术，有12.5%的卵母细胞受精并发育到PN期。4.在人精子和仓鼠卵母细胞的试验中，有3.0%的仓鼠卵母细胞在相同的电融合条件下受精。尽管电融合的成功率很低，最佳的电融合条件还需进一步研究，但我们的初步结果为精子功能障碍患者的治疗提供了重要的前景。