CAJ | 학술논문

目的：对Wistar大鼠骨髓源神经干细胞进行分离、培养和鉴定，观察其生长方式和分化特征。方法：运用无血清培养基对Wistar大鼠骨髓基质细胞进行培养，对分离获得的悬浮生长的神经球采用免疫组织化学法检测CD133和Nestin表达情况。用血清诱导其分化，分化7 d后，采用免疫荧光细胞化学染色方法检测分化后GFAP、Map2、β-tubulinⅢ、Galc的表达情况。结果：大鼠骨髓基质细胞在无血清培养基中，呈悬浮状态生长，形成细胞球，经免疫荧光检测，细胞球呈CD133和Nestin阳性。将细胞球转入含血清培养基后，转为贴壁生长，经免疫荧光检测大部分已分化细胞呈GFAP阳性，少部分细胞呈MAP-2、β-tubulinⅢ及Galc阳性。结论：采用含bFGF、EGF的无血清培养基可培养出呈球状聚集生长、具多向分化潜能的大鼠骨髓源神经干细胞。
목적：대Wistar대서골수원신경간세포진행분리、배양화감정，관찰기생장방식화분화특정。방법：운용무혈청배양기대Wistar대서골수기질세포진행배양，대분리획득적현부생장적신경구채용면역조직화학법검측CD133화Nestin표체정황。용혈청유도기분화，분화7 d후，채용면역형광세포화학염색방법검측분화후GFAP、Map2、β-tubulinⅢ、Galc적표체정황。결과：대서골수기질세포재무혈청배양기중，정현부상태생장，형성세포구，경면역형광검측，세포구정CD133화Nestin양성。장세포구전입함혈청배양기후，전위첩벽생장，경면역형광검측대부분이분화세포정GFAP양성，소부분세포정MAP-2、β-tubulinⅢ급Galc양성。결론：채용함bFGF、EGF적무혈청배양기가배양출정구상취집생장、구다향분화잠능적대서골수원신경간세포。
Objective:To isolate,culture and identify the bone marrow mesenchymal stem cells-derived neural stem cells(BMSCs-NSCs)in order to observe their growth pattern and differentiation features.Method:Bone marrow mesenchymal stem cells of the Wistar rats were cultured in serum-free medium.Floating neural spheres were formed in serum-free medium,the expression of CD133 and Nestin in the neural spheres was tested by immunohistochemistry. Then floating neural spheres were transferred into serum containing medium for induction differentiation,the expression of GFAP,Map2,β-tubulinⅢand Galc were detected by immunocytochemistry 7 days later.Result:Bone marrow mesenchymal stem cells lived in serum-free medium‘floating’and it formed neural sphere.CD133 and Nestin were positive in neural sphere cells according to the results of immunohistochemistry.When these neural sphere cells were transferred into serum-containing medium,they grew adhering to the wall.GFAPMAP-2,β-tubulinⅢand Galc were positive in the cells differentiated from neural spheres.Conclusion:Bone marrow mesenchymal stem cells-derived neural stem cells which growing like a ball and having multipotential differentiation are successfully cultured in serum-free medium with bFGF and EGF.