3种培养体系下人胚胎干细胞神经诱导分化的比较  

Comparison of neural induction of human embryonic stem cells in three culture systems

作　　者：袁丁[1,2] 林戈[1,2] 卢光琇[1,2] 

机构地区：[1]人类干细胞国家工程研究中心,湖南长沙410078 [2]中南大学生殖与干细胞工程研究所,湖南长沙410078

出　　处：《西北农林科技大学学报（自然科学版）》2010年第4期31-36,共6页Journal of Northwest A&F University(Natural Science Edition)

基　　金：国家"863"计划项目(2006AA02A102);国家"973"计划项目(2007CB948103)

摘　　要：【目的】比较人胚胎干细胞在人胚胎成纤维细胞、鼠胚胎成纤维细胞饲养层和无饲养层3个诱导体系下形成神经上皮祖细胞的能力。【方法】将人胚胎干细胞在人、鼠胚胎成纤维细胞饲养层或无饲养层体系中诱导10 d后,检测其胚胎干细胞和神经上皮祖细胞相关基因(Oct4、Nestin、Pax6、Sox1、Sox2、Sox3、β-actin)的表达,用免疫荧光染色法检测神经上皮祖细胞抗原标记Pax6、Musashi、Nestin和胚胎干细胞抗原标记SSEA4的表达。【结果】3种诱导体系下均有神经上皮祖细胞形成,均表达神经上皮祖细胞抗原标记Pax6、Musashi、Nestin及Pax6、Nes-tin、Sox家族基因。无饲养层体系诱导后的细胞,Oct4基因的表达高于有饲养层体系;其SSEA4染色阳性细胞比例为(26.3±3.5)%,高于有饲养层体系;其Pax6和Musashi染色阳性细胞比例分别为(81.0±3.0)%,(79.0±4.4)%,均低于有饲养层细胞体系。【结论】人胚胎干细胞在有饲养层和无饲养层体系中均可诱导分化形成神经上皮祖细胞,但无饲养层体系的效率相对较低,且有未分化胚胎干细胞残余。【Objective】 The study was done to compare the induction efficiency of human embryonic stem cells to neuroepithelial progenitor cells in human embryonic fibroblasts,mouse embryonic fibroblasts and feeder free systems.【Method】 Human embryonic stem cells cultured on human or mouse feeder cells or in feeder free conditions were induced into neuroepithelial progenitor cells,and 10 days later embryonic stem cell related genes and neuroepithelial progenitor related genes were tested,so were immunofluorescence cytochemistry of neuroepithelial progenitor markers Pax6,Musashi and Nestin and embryonic stem cell marker SSEA4.【Result】 Neuroepithelial progenitors could be induced in three inducing systems,expressing neuroepithelial progenitor antigen markers Pax6,Musashi and Nestin as well as Pax6,Nestin and Sox family genes.The expression of Oct4 in feeder free system was higher than that in feeder containing systems,so was SSEA4 positive cell percentage,which was （26.3±3.5）%,while Pax6 and Musashi positive cell percentage was lower than that in feeder containing systems,which was （81.0±3.0）% and （79.0±4.4）% respectively.【Conclusion】 Human embryonic stem cells can differentiate into neuroepithelial progenitor cells in both feeder containing and feeder free systems,yet feeder free system has a relatively lower inductive efficiency with part of undifferentiated embryonic stem cells.

关 键 词：胚胎干细胞 神经诱导 饲养层细胞 

分 类 号：Q813.1[生物学—生物工程]