The basic study of cryopreservation and in vitro maturation of encapsulated human immature follicles

封装的人未成熟卵泡的冷冻保存和体外成熟的基础研究

基本信息

批准号：
15591725
负责人：
SENGOKU Kazuo
金额：
$ 2.18万
依托单位：
Asahikawa Medical College
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15591725/
关键词：
immature follicle microcapsule cryopreservation in vitro culture spermatogenesis aging 卵子活性化体外成熟

项目摘要

We studied the possibility of cryopreservation and in vitro maturation of immature human ovarian follicles using microcapsule methods.In addition, we investigated the mechanisms of follicular development by analyzing the oocyte factor.The following results were obtained.1.Human unfertilized oocytes were successfully cryopreserved with vitrification methods (10% EG and 15% EG+15% DMSO+0.5M sucrose). However, in vitro growth of encapsulated immature follicles were not obtained. The expression of GDF-9,CD9,CD98, C-kit mRNA were confirmed in human un fertilized oocytes. These results may suggest the possible future application of this methods for preservation immature follicles2.The dynamic and cell-specific expression pattern of HB-EGF observed in our study suggests that HB-EGF may be potentially important local regulator of follicular development.3.Our results regarding sperm-oocyte interaction support the hypothesis that one of the sperm oolemmal binding mechanism can be inhibited by integrin antibodies but that this mechanism does not play an essential role in the binding and fusion process4.Our studies regarding the genetic mechanisms implicate as a cause of male infertility suggest that OPPO1 and MEI1 may play a role in human spermatogenesis.5.We investigated the nuclear kinetics of oocyte activation of aged human oocytes following combined treatment with calcium ionophore and puromycin. It is concluded that the nuclear kinetics of parthenogenetically activated human oocytes appears to be more rapid in in vitro aging oocytes.Although successful in vitro maturation was not obtained using encapsulated human immature follicles, our preliminary results may offer important aspects of understanding the mechanisms of human follicular development.

我们研究了使用微胶囊方法研究冷冻保存和体外成熟的可能性。此外，我们通过分析卵母细胞因子研究了卵泡发育的机制。获得以下结果。获得以下结果。1.Human未经使用的卵母细胞成功地使用了5％（10％ED）（10％EG）（5％）（10％ED）。蔗糖）。但是，未获得封装未成熟卵泡的体外生长。在人类联合受精的卵母细胞中证实了GDF-9，CD9，CD98，C-KIT mRNA的表达。这些结果可能表明，这种方法可能将来可能应用这种方法用于保存不成熟的卵泡。在我们的研究中观察到的HB-EGF的动态和细胞特异性表达模式表明，HB-EGF可能是卵泡发育的潜在重要的局部调节剂。3。您对精子 - 元素相互作用的结果可能是通过侵害了精子的方法，而这些方法可以抑制了这种方法，而这些机制是侵蚀性的。在结合和融合过程中起着至关重要的作用。您关于遗传机制的研究所暗示是男性不育症的原因，这表明Oppo1和MEI1可能在人类的精子发生中起作用。5。我们研究了衰老人类卵母细胞的核能激活型卵母细胞的核动力学，该动力学与造成的卵巢粒子和卵巢粒子综合治疗后。得出的结论是，在体外衰老的卵母细胞中，孤og活化的人卵母细胞的核动力学似乎更快。尽管未使用包裹的人类不成熟的卵泡成功获得体外成熟，但我们的初步结果可能提供了理解人类卵泡发展机制的重要方面。