Study on the selection of normal sperm and the method of fusion of sperm and oocytes for micro-feritlization.

发明了一种新的精子选择方法--游动移位(SS)法。与传统的上游(SU)方法相比，该方法回收的活动精子更多。PSA柱法选择性地提取形态正常的精子。SS法和PSA柱法联合应用时，精子密度和精子形态百分率均高于SU法，而精子畸形率与SU法相似。由于蔗糖对卵母细胞有毒性，对几种精子亚区受精(Suzi)方法进行了改进，使金黄地鼠卵母细胞受精率最低(%D)，最高受精率(%F)，但不使用蔗糖扩大亚区空间。用5 MU的钙离子载体对精子进行预处理，可提高去透明带仓鼠卵母细胞的%F。采用改良苏子法治疗严重男性不育，与常规苏子法相比，胚胎卵裂率和卵裂率均有显著提高，但妊娠率无显著差异。当精子滴中精子浓度稀释100倍至每毫升10万个精子时，金黄地鼠卵母细胞卵胞浆内单精子注射(ICSI)的卵裂率和卵裂率显著提高。建议在ICSI前切开精子中段增加%F。使用压电操作器可显著降低%D和增加%F。对严重不育症患者采用稀释精子密度、切开精子中段、使用压电针等改进的ICSI方法，受精卵、卵裂胚胎数及每周期妊娠率均较常规ICSI方法显著增加，每周期妊娠率达20%。

The swim-shift (SS) methoda novel method of sperm selection, was invented. It retrieved more motile sperm than the conventional swim-up (SU) method. The PSA-column method retrieved morphologically normal sperm selectively. When SS method and PSA-column method were combined, a sperm concentration and %norma morphology were higher and %mortility was similar, compared with the SU method alone. Several procedures of subzonal insemination of sperm (SUZI) were modified to give a minimal %damage (%D) and maximal %fertilization (%F) of hamster oocytes with human sperm without use of sucrose for expansion of subzonal space, becaouse sucrose toxicity to oocytes was suggested. Pretreatment of sperm with 5 muM calcium ionphore improved %F of zona-free hamster oocytes. When the improved procedures of SUZI was applied for treatments of severe male infertility, %F and %embryo cleavage were improved significantly but %pregnancy was not, compared with the conventional procedures of SUZI.When sperm concentration of sperm droplet was diluted by 100 times to give 100,000 per ml, %F of hamster oocytes with human sperm by intracytoplasmic sperm injection (ICSI) was significantly increased. It was suggested that incision of sperm midpiece just before ICSI increased %F.A use of Piezo-manipulator decreased %D and increased %F significantly. When the improved procedures of ICSI with dilution of sperm concentration, incision of sperm midpiece, and use of Piezo-manipulator was applied for treatments of severe male infertility, numbers of fertilized oocytes and cleaved embryos and %pregnancy per cycle were significantly increased compared with conventional procedures of ICSI,resulting in 20% of ongoing pregnancy rate per cycle.