CAJ | 학술논문

　　背景：目前神经干细胞多由动物获得，不适合人类临床移植治疗。目的：探索体外环境下人胚胎纹状体来源神经干细胞的培养方法，同时观察其生物学特性。方法：取经水囊引产的孕8-16周人胚胎纹状体，体外用无血清 DMEM 培养基进行培养，待细胞形成神经球后进行传代，并应用含体积分数10%胎牛血清的 DMEM/F12培养液进行诱导分化。结果与结论：体外培养的人胚胎纹状体来源神经干细胞生长迅速，表达神经干细胞标志物 nestin。克隆形成实验显示细胞克隆形成率为6.0%-7.0%；BrdU 掺入实验显示细胞增殖率为37.9%。免疫荧光染色显示经诱导分化的细胞表达神经元标志物Ⅲ型β微管蛋白、星形胶质细胞标志物胶质纤维酸性蛋白及神经干细胞标志物nestin，但不表达少突胶质细胞标志物髓鞘碱性蛋白。可见人胚胎纹状体来源神经干细胞在体外无血清条件下可保持其生物学特点，具有自我更新能力，经胎牛血清诱导后可向神经元及星形胶质细胞分化。
　　배경：목전신경간세포다유동물획득，불괄합인류림상이식치료。목적：탐색체외배경하인배태문상체래원신경간세포적배양방법，동시관찰기생물학특성。방법：취경수낭인산적잉8-16주인배태문상체，체외용무혈청 DMEM 배양기진행배양，대세포형성신경구후진행전대，병응용함체적분수10%태우혈청적 DMEM/F12배양액진행유도분화。결과여결론：체외배양적인배태문상체래원신경간세포생장신속，표체신경간세포표지물 nestin。극륭형성실험현시세포극륭형성솔위6.0%-7.0%；BrdU 참입실험현시세포증식솔위37.9%。면역형광염색현시경유도분화적세포표체신경원표지물Ⅲ형β미관단백、성형효질세포표지물효질섬유산성단백급신경간세포표지물nestin，단불표체소돌효질세포표지물수초감성단백。가견인배태문상체래원신경간세포재체외무혈청조건하가보지기생물학특점，구유자아경신능력，경태우혈청유도후가향신경원급성형효질세포분화。
BACKGROUND: Neural stem cells are always derived from animals, and unsuitable for human transplantation treatment. OBJECTIVE: To explore the in vitro culture methods of human embryonic striatum-derived neural stem cells, and to observe the biological characteristics. METHODS: The human embryonic striatum were separated from the embryo at a gestational age of 8-16 weeks that received induction of labor with water bag, and then the embryonic striatum was in vitro cultured in the serum-free Dulbecco’s modified Eagle’s medium. The cells were passaged after neurospheres formation, and then the cells were induced to differentiation with the Dulbecco’s modified Eagle’s medium/F12 containing 10% fetal bovine serum. RESULTS AND CONCLUSION: The in vitro cultured human embryonic striatum-derived neural stem cells grew rapidly and could express nestin. Colony formation assay showed the cel clone formation rate was 6.0%-7.0%. 5-Bromodeoxyuridine incorporation assay showed the cel proliferation rate was 37.9%. Immunofluorescence staining showed that the cells after induction and differentiation could express Tuj-1, glial fibril ary acidic protein and nestin, but not express myelin basic protein. The results indicate that human embryonic striatum-derived neural stem cells cultured in the serum-free medium can maintain their biological characteristics and have self-renewal capacity, and the cells can differentiate into the neurons and astrocytes induced by the fetal bovine serum.