细胞筛网作为载体运用于玻璃化法冻存人卵巢组织的效果分析
作者:
作者单位:

作者简介:

通讯作者:

中图分类号:

基金项目:

国家973课题(2009CB941701);江苏省临床医学科技专项(BL2012009);江苏省临床生殖医学中心(ZX201110)


Efficiency of cell strainer-based vitrification in human ovarian tissue cryopreservation
Author:
Affiliation:

Fund Project:

  • 摘要
  • |
  • 图/表
  • |
  • 访问统计
  • |
  • 参考文献
  • |
  • 相似文献
  • |
  • 引证文献
  • |
  • 资源附件
  • |
  • 文章评论
    摘要:

    目的:研究细胞筛网作为载体运用于玻璃化冷冻人卵巢组织的效果,并寻找合适的冷冻剂浓度。方法:选择来自卵巢切除-子宫内膜异位-卵巢浆液性囊肿患者的卵巢组织共10例,以细胞筛网为载体,以二甲基亚砜-乙二醇和蔗糖为冷冻保护剂进行玻璃化冷冻。分别以二甲基亚砜和乙二醇等浓度混配(12%-15%-18%和20%)和0.5 mol/L的蔗糖配成4组不同的冷冻液,并以新鲜组织及常规程序化冷冻组织为对照,观察不同冷冻液组合对卵巢组织始基卵泡的形态学-凋亡以及体外培养后雌激素-孕酮和乳酸脱氢酶的影响,评价冷冻效果。结果:与新鲜组相比,细胞网筛玻璃化冷冻的卵巢组织冻融后,始基卵泡正常率虽有所降低,但优于常规程序冷冻组,其中18%组和15%组的正常率显著增高。组织细胞的凋亡率比新鲜组有所增加,但低于常规程序冷冻组,其中15%组和12%组明显降低。因此,15%组的冷冻效果最好。比较了该浓度组合对卵巢组织培养后的雌激素-孕酮-乳酸脱氢酶的影响,结果发现,与新鲜组和程序化冷冻组相比,培养液中雌激素水平无明显差异,孕酮水平略好于程序组,但乳酸脱氢酶与新鲜组的变化完全一样,都显著优于程序化组。结论:细胞筛网玻璃化冷冻法优于程序化冷冻法,可作为临床卵巢组织冷冻的一种方法,15%的二甲基亚砜和乙二醇等浓度组合效果最佳。

    Abstract:

    Objective:To investigate the efficiency of cell strainer-based vitrification in human ovarian tissue cryopreservation,and to find suitable concentration of cryoprotectants. Methods:Human ovarian biopsy tissues were obtained from ten female patients undergoing oophorectomy,endometriosis surgery, and ovarian cystectomy. The ovarian tissues were vitrificated by cryoprotectants containing ethylene glycol (EG),dimethylsulfoxide (DMSO) and sucrose using cell strainer as a device. The cryoprotectant mixtures were formulated as follows:12%DMSO + 12%EG + 0.5 mol/L sucrose,15%DMSO + 15%EG + 0.5 mol/L sucrose,18%DMSO + 18%EG + 0.5 mol/L sucrose,20%DMSO +20%EG + 0.5 mol/L sucrose. The morphology of the primordial follicles,cellular apoptosis in ovarian tissues,estradiol(E2),progesterone(P) and lactate dehydrogenase (LDH) in culture medium were evaluated. The fresh tissues and the tissues in programmed cryopreservation were used as controls. Results:Compared with the fresh group,the proportion of normal primordial follicle declined a little in the cell strainer-based vitrification group but was higher than the programmed cryopreservation group,of which the proportion increased significantly in the 18% and 15% groups. Furthermore,the proportion of TUNEL-positive primordial follicles in the vitrification group was higher than that of the fresh group but was lower than that of the programmed cryopreservation group,of which the proportion decreased significantly in the 15% and 12% groups. Therefore,15% cryoprotectant mixtures was optimum for the ovarian tissues preservation. After recovery culturing the ovarian tissues from vitrification in 15% cryoprotectant mixtures,the levels of E2 and P were similar to the controls. However,the level of LDH was almost same as the fresh group,both of which were much more superior than the programmed cryopreservation group. Conclusion:The cell strainer-based vitrification is superior than the programmed cryopreservation,it could be a new vitrification method for clinical human ovarian tissue cryopreservation. The mixture containing 15%DMSO + 15%EG + 0.5 mol/L sucrose should be a optimized cryoprotectant.

    参考文献
    相似文献
    引证文献
引用本文

马 龙,蔡令波,严正杰,冯 婷,董 娟,刘嘉茵,王守林.细胞筛网作为载体运用于玻璃化法冻存人卵巢组织的效果分析[J].南京医科大学学报(自然科学版),2013,(9):1180-1185

复制
分享
文章指标
  • 点击次数:
  • 下载次数:
  • HTML阅读次数:
  • 引用次数:
历史
  • 收稿日期:2013-05-03
  • 最后修改日期:
  • 录用日期:
  • 在线发布日期: 2013-09-18
  • 出版日期:
通知关闭
郑重声明