GM-1对人胚胎干细胞定向诱导分化神经细胞的作用  

Role of GM-1 in human embryonic stem cells differentiating into neural cells

作　　者：高选[1,2] 颜军昊[2] 宦晴[2] 黄色新[2] 徐凯[2] 陈子江[2] 王磊[1] 

机构地区：[1]山东大学药学院药物分析研究所,济南250012 [2]山东大学附属省立医院生殖医学中心山东省生殖医学重点实验室,济南250021

出　　处：《山东大学学报（医学版）》2009年第12期21-24,共4页Journal of Shandong University:Health Sciences

基　　金：国家重点基础研究发展计划973项目(2006CB944004);山东省自然科学基金重点项目(Z2002C05);山东省科技发展重点资助项目(2002DD1DBA1)

摘　　要：目的探索外源性神经节苷脂(GM-1)对人胚胎干细胞(hESC)诱导分化成神经细胞的作用,为临床治疗提供理论依据。方法将人胚胎干细胞(SDU-hESm-1)悬浮培养形成拟胚体(EB),EB再贴壁培养诱导分化神经细胞,分别于悬浮培养EB阶段和EB贴壁培养阶段的神经分化培养基中加入浓度为0、0.5、5、50μg/mL的GM-1,观察EB的形成,计数神经标记性抗原Nestin和β-Tubulin阳性细胞百分率。结果含GM-1的神经分化培养基诱导生成"透亮EB",分化出Nestin和β-Tubulin阳性细胞明显多于不含GM-1培养基诱导出的神经细胞数量,不含GM-1形成的EB发黑;在悬浮培养形成EB开始加GM-1比EB贴壁培养开始加GM-1产生Nestin和β-Tubulin阳性细胞数目更多,但2个加GM-1阶段产生的神经细胞百分率均随GM-1浓度增加而增大。结论首次发现GM-1可以诱导hESC定向分化神经细胞,定向产生神经前体细胞(Nestin阳性),在hESC定向分化神经细胞的过程中,悬浮培养EB阶段加入GM-1更有利于hESC的定向分化。Objective To explore the possible role of ganglioside GM-1 in human embryonic stem cells differentiating into neural cells and to provide a method for clinical treatment in the future.Methods Human embryonic stem cells underwent suspension culture to form embryonic bodies（EB）and then the EB underwent adherent culture to induce neural cells.0,0.5 μg/ml,5 μg/ml,and 50 μg/ml GM-1 were respectively added to the neural differentiation culture media and then morphology of the EB and neural cells were observed,and positive cells of Nestin and β-Tubulin were counted.Results Light EB were formed when culture media were added to GM-1,while dark EB were formed when culture media was not added to GM-1.Nestin and β-Tubulin positive cells induced by neural differentiation culture media with GM-1 were more significant than those without GM-1.There were more Nestin and β-Tubulin positive cells when adding GM-1 in suspension culture than in adherent culture,also with an increase of the concentration of GM-1,the Nestin and β-Tubulin positive cells increased in both media.Conclusion GM-1 may directionally induce hESCs into neural progenitor cells.GM-1 added in suspension culture media is helpful in the directional differentiation of hESCs.

关 键 词：干细胞 G(M1)神经节苷脂 细胞 培养的 

分 类 号：R687.34[医药卫生—骨科学] R329.2[医药卫生—外科学]