本文關鍵詞： 卵巢 細胞 培養(yǎng)的 受精 體外 組織構建 組織工程 卵巢顆粒細胞 裂解法 沉淀法 密度梯度離心法 Ⅰ型膠原酶 透明質(zhì)酸酶 自體卵泡液　出處：《中國組織工程研究》2015年46期 　論文類型：期刊論文

【摘要】：背景:建立分離培養(yǎng)顆粒細胞快速有效的方法也是提高體外受精-胚胎移植成功率關鍵的一步。雖然目前文獻中有較多關于人卵巢顆粒細胞分離方法的報道,但在細胞數(shù)量、純度及后續(xù)生長等方面不盡人意。目的:建立有效的分離提純、體外培養(yǎng)人卵巢黃素化顆粒細胞的方法。方法:收集體外受精-胚胎移植取卵時的卵泡液,用裂解法、沉淀法、密度梯度離心法分離,Ⅰ型膠原酶或透明質(zhì)酸酶消化顆粒細胞黏液團并接種在培養(yǎng)皿中進行培養(yǎng),培養(yǎng)液中加入或不加自體卵泡液。結果與結論:用裂解法得到的顆粒細胞數(shù)較沉淀法和密度梯度離心法得到的細胞數(shù)多(P0.05,P0.05);3種方法提取的顆粒細胞活性比較無明顯差異;培養(yǎng)24 h后沉淀法貼壁細胞數(shù)最多(P0.05),而密度梯度離心法貼壁細胞數(shù)最少(P0.05);透明質(zhì)酸酶消化顆粒細胞較Ⅰ型膠原酶耗時少且消化徹底;加入自體卵泡液能夠促進顆粒細胞生長和存活。結果證實,沉淀法提取顆粒細胞雖然耗時長,但培養(yǎng)的細胞存活率高、培養(yǎng)后收獲的細胞數(shù)多;透明質(zhì)酸酶較Ⅰ型膠原酶更適合消化顆粒細胞黏液團;在培養(yǎng)液中加入自體卵泡液更有益于顆粒細胞生長。
[Abstract]:Background: to establish a rapid and effective method for isolation and culture of granulosa cells is also a key step to improve the success rate of in vitro fertilization and embryo transfer. Objective: to establish an effective method for isolation and purification of human ovarian luteinized granulosa cells in vitro. By density gradient centrifugation, granulosa cell mucus was digested by type I collagenase or hyaluronidase and cultured in culture dish. Results and conclusion: the number of granulosa cells obtained by cleavage method was more than that by precipitation method and density gradient centrifugation method. After 24 h culture, the number of adherent cells by precipitation method was the highest, and the number of adherent cells by density gradient centrifugation was the least, and hyaluronidase digestion of granulosa cells took less time and digested thoroughly than that of type 鈪，

本文編號：1510416