發(fā)布時(shí)間：2018-02-14 09:43

  本文關(guān)鍵詞： 卵巢 細(xì)胞 培養(yǎng)的 受精 體外 組織構(gòu)建 組織工程 卵巢顆粒細(xì)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(guó)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(xì)胞快速有效的方法也是提高體外受精-胚胎移植成功率關(guān)鍵的一步。雖然目前文獻(xiàn)中有較多關(guān)于人卵巢顆粒細(xì)胞分離方法的報(bào)道,但在細(xì)胞數(shù)量、純度及后續(xù)生長(zhǎng)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(xì)胞的方法。方法:收集體外受精-胚胎移植取卵時(shí)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(xì)胞黏液團(tuán)并接種在培養(yǎng)皿中進(jìn)行培養(yǎng),培養(yǎng)液中加入或不加自體卵泡液。結(jié)果與結(jié)論:用裂解法得到的顆粒細(xì)胞數(shù)較沉淀法和密度梯度離心法得到的細(xì)胞數(shù)多(P0.05,P0.05);3種方法提取的顆粒細(xì)胞活性比較無明顯差異;培養(yǎng)24 h后沉淀法貼壁細(xì)胞數(shù)最多(P0.05),而密度梯度離心法貼壁細(xì)胞數(shù)最少(P0.05);透明質(zhì)酸酶消化顆粒細(xì)胞較Ⅰ型膠原酶耗時(shí)少且消化徹底;加入自體卵泡液能夠促進(jìn)顆粒細(xì)胞生長(zhǎng)和存活。結(jié)果證實(shí),沉淀法提取顆粒細(xì)胞雖然耗時(shí)長(zhǎng),但培養(yǎng)的細(xì)胞存活率高、培養(yǎng)后收獲的細(xì)胞數(shù)多;透明質(zhì)酸酶較Ⅰ型膠原酶更適合消化顆粒細(xì)胞黏液團(tuán);在培養(yǎng)液中加入自體卵泡液更有益于顆粒細(xì)胞生長(zhǎng)。
[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(hào)：1510416