CAJ | 학술논문

目的：探讨不同胎龄胎鼠脊髓源性神经干细胞(NSCs)的增殖分化能力。方法 Sprague-Dawley孕鼠分为A组(孕12 d)、B组(孕14 d)、C组(孕16 d)，酶消化法结合机械分离法提取细胞，检测不同时间各组神经干细胞球的直径、数量，CCK-8法绘制细胞生长曲线。BrdU/Nestin免疫荧光染色鉴定NSCs。10%胎牛血清诱导分化后行β-tubulinⅢ、胶质纤维酸性蛋白(GFAP)免疫荧光染色，比较各组神经元的分化比例。结果各组细胞BrdU、Nestin、β-tubulinⅢ、GFAP免疫荧光染色均为阳性。B组β-tubulinⅢ染色阳性的比例最高；B、C两组各时间点神经球直径、数量及神经元的分化比例高于A组(P<0.05)。第3代NSCs培养第5天， B组细胞吸光度高于C组(P<0.05)。结论孕14 d左右取材培养的脊髓神经干细胞生物学特性稳定，增殖活力强，诱导分化后神经元的分化比例较高。
목적：탐토불동태령태서척수원성신경간세포(NSCs)적증식분화능력。방법 Sprague-Dawley잉서분위A조(잉12 d)、B조(잉14 d)、C조(잉16 d)，매소화법결합궤계분리법제취세포，검측불동시간각조신경간세포구적직경、수량，CCK-8법회제세포생장곡선。BrdU/Nestin면역형광염색감정NSCs。10%태우혈청유도분화후행β-tubulinⅢ、효질섬유산성단백(GFAP)면역형광염색，비교각조신경원적분화비례。결과각조세포BrdU、Nestin、β-tubulinⅢ、GFAP면역형광염색균위양성。B조β-tubulinⅢ염색양성적비례최고；B、C량조각시간점신경구직경、수량급신경원적분화비례고우A조(P<0.05)。제3대NSCs배양제5천， B조세포흡광도고우C조(P<0.05)。결론잉14 d좌우취재배양적척수신경간세포생물학특성은정，증식활력강，유도분화후신경원적분화비례교고。
Objective To study the differentiation and proliferation ability of the spinal neural stem cells (NSCs) at different gestational ages in fetal rats. Methods Sprague-Dawley fetal rats were divided into group A (12 days of pregnancy), group B (14 days of pregnancy) and group C (16 days of pregnancy). NSCs were separated with enzyme-assisted microdissection. The diameter and numbers of NSCs balls were measured at different time. The cell growth curve was drawn with CCK8 colorimeter. NSCs were identified with BrdU/Nestin immuno-histochemical staining. They were induced with 10%fetal bovine serum for 10 days, and the expression ofβ-tubulinⅢand glial fibrillary acidic protein was detected with immunocytochemistry. Results There were cells expressed BrdU, Nestin,β-tubulinⅢand GFAP in all the group. The most cells (22.74±0.79%) expressedβ-tubulinⅢin the group B, but no significant difference between group B and group C. The cell vitality on the 5th day of third-generation neural stem cells was the most in group B. Conclusion For enzyme-assisted microdissection, it may obtain more neurons to isolate the neural stem cells from 14 days of pregnancy pregnant rats.