序贯培养法诱导小鼠胚胎干细胞向神经细胞分化的研究  

The neural induction of the mouse embryonic stem cells by the sequential culture method

作　　者：顾文佳[1] 李宏图[1,2] 刘晓玉[1] 庞希宁[1] 

机构地区：[1]中国医科大学发育生物学教研室卫生部细胞生物学重点实验室,辽宁沈阳110001 [2]辽宁省计划生育科学研究院辽宁省生殖医学重点实验室,辽宁沈阳110031

出　　处：《解剖科学进展》2009年第4期370-375,共6页Progress of Anatomical Sciences

基　　金：辽宁省自然科学基金资助项目(20062090);辽宁省教育厅资助项目(2004D227)

摘　　要：目的诱导小鼠胚胎干细胞向神经细胞分化。方法通过"序贯培养法"诱导小鼠胚胎干细胞分化为神经细胞,并应用RT-PCR、免疫荧光、端粒酶活性及基因芯片等对诱导结果进行检测。结果分化细胞表达神经元特异性烯醇化酶(NSE)、低分子量神经微丝蛋白(NF-L)、神经细胞粘附分子1(NCAM1)、微管相关蛋白Tau等多种神经元标志物,免疫荧光显示NSE阳性率为(81±9.2)%,而神经胶质细胞原纤维酸性蛋白(GFAP)几乎不表达。基因芯片分析结果显示,在3张芯片中有8179个基因共表达,另有998个基因的表达完全不同,且有多种神经元基因的标志物表达明显升高。结论通过"序贯培养法"能有效地诱导小鼠胚胎干细胞向神经元分化。Objective To induce the differentiation of mouse embryonic stem cells（ESCs）into neural cells in vitro. Methods Through the sequential culture method, mouse ESCs were differentiated into neural cells that were detected by RT-PCR, immunofluorescence, telomerase activity and genechip. Results The differentiated cells were immunopositive for many neural markers, such as NSE （neuron specific enolase）, NF-L （neurofilament triplet L）, NCAM1 （neural cell adhesion moleculel） and Tau, and （81 ± 9.2）% induced cells were NSE-positive. Few glial fibrillary acidic protein （GFAP）-positive cells were detected. Genechip analysis showed that 8179 genes were co-expressed and 998 genes were significantly differentially expressed in three genechips and expression of many neuronal marker genes was significantly increased. Conclusion The sequential culture method can efficiently induce the differentiation of mouse ESCs into neurons in vitro.

关 键 词：胚胎干细胞 神经细胞 序贯培养法 诱导分化 小鼠 

分 类 号：Q254[生物学—细胞生物学]