大鼠BMSCs条件培养基对NSCs形态生长及分化的影响.PDF

网络出版时间：2015-08-21 10:43:33 网络出版地址：/kcms/detail/31.2035.Q1043.002.html 中国细胞生物学学报 Chinese Journal of Cell Biology 2015, 37(8):1102-1108 DOI: 10.11844/cj cb.2015.08.0082 大鼠BMSCs条件培养基对NSCs形态、生长及 分化的影响 纪志华 钟贞浩 史方富 唐崧杰 孟志斌* (海南医学院附属医院骨科, 海口570102) 摘要 通过对SD大鼠骨髓间充质干细胞(bone mesenchymal stem cells, BMSCs)及新生鼠神经 干细胞(neural stem cells, NSCs)进行体外分离、培养及鉴定后, 观察BMSCs条件培养基对NSCs向神 经细胞分化的影响。采用全贴壁培养法培养BMSCs, 并采用流式细胞术鉴定其特异性表面抗原标记。 无血清技术培养NSCs, 采用免疫荧光技术鉴定其特异性抗原标记。BMSCs条件培养基(含10%胎牛 血清的DMEM培养基)对NSCs诱导7 d后, 镜下观察细胞形态和生长, 并采用免疫荧光技术检测NSCs 分化。BMSCs高表达CD90、CD29(90%), 而CD45呈阴性表达。免疫荧光染色显示, NSCs标记蛋白 Nestin 、SOX2为阳性。NSCs经BMSCs培养液诱导分化7 d后经免疫荧光鉴定, MAP-2及GFAP呈阳性, 阳性率分别为73.80%和50.47%; NSCs经胎牛血清(fetal bovine serum, FBS)诱导分化7 d后经免疫荧光 鉴定, MAP-2及GFAP呈阳性, 阳性率分别为42.14%和31.90%。BMSCs条件培养基可诱导NSCs分化 为神经元及星形胶质细胞, 其中BMSCs分泌的细胞因子在诱导分化中可能发挥重要作用。 关键词 骨髓间充质干细胞; 神经干细胞; 培养; 诱导; 分化; 鉴定; 免疫细胞荧光染色 The Effect of Bone Mesenchymal Stem Cells Culture Medium on the Morphology, Growth and Differentiation of Neural Stem Cells Ji Zhihua, Zhong Zhenhao, Shi Fangfu, Tang Songjie, Meng Zhibin* (Department of Orthopaedics, Affiliated Hospital of Hainan Medical University, Haikou 570102, China) Abstract To observe the effect of BMSCs (bone mesenchymal stem cells) conditioned medium on the differentiation of NSCs (neural stem cells) after the isolation, culture and identification of SD rat BMSCs and NSCs. BMSCs were cultured using method of all-adherent culture and identified with specific antigen marker by flow cytometry. NSCs were cultured us