摘要
目的探索传代对胚胎神经干细胞的增生、分化和凋亡等生物学性状的影响。方法原代培养后(P0)和经过两次传代后(P2)的神经干细胞用BrdU细胞增生检测试剂盒检测增生能力;用Annexin V-FITC凋亡试剂盒检测早期凋亡率;诱导分化7天后,分别进行MAP2和GFAP免疫荧光标记和Hoechst 33342核染色,计算MAP2+细胞和GFAP+细胞占Hoechst+细胞的各自比例。结果P0神经干细胞的OD值是:1.1899±0.0485(n=13),P2为0.3526±0.1053(n=11,P=0.000);P0的早期凋亡率为40.91%±12.74%(n=8),P2为24.31%±13.01%(n=9,P=0.018);P0和P2的神经元细胞比例分别为:58.81%±9.69%(n=10)和27.84%±5.70%(n=10,P=0.000),星形胶质细胞比例分别为22.85%±4.36%(n=10)和49.32%±6.95%(n=10,P=0.000)。结论传代后的神经干细胞的增生能力和早期凋亡率都会变弱,分化为神经元细胞的比例会降低,而星形胶质细胞的比例增大。
Objective To investigate the effects of passaging on the proliferation, differentiation and apoptosis of neural stem cells. Methods Neural stem cells both from prime culture （PO） and twice - passaged culture （ P2 ） were tested by enzyme immunoassay for proliferation and fluorescent activated cell sorting for apoptosis by Annexin V - FITC and propjdium iodide. After being enduced to differentiation for 7 days, they were detected by the neuronal marker MAP2 and the astrocyte marker GFAP with nuclei staining by Hoechst 33342 additionally. The ratios of MAP2 ^＋/Hoechst ^＋ and GFAP ^＋/Hoechst ^＋ were calculated. Results The absorbance of NSCs from PO and P2 is 1.1899 ± 0. 0485 （ n = 13 ） and 0. 3526 ± 0. 1053 （ n = 11 ） （ P = 0.000）. Analyses of apoptosis using cytoflurometry are 40.91% ± 12. 74% forPO（n=8）and24.31% ±13.01% for P2（n=9,P=0.018）. The ratios ofMAP2^＋/Hoechst^＋ and GFAP＋/Hoechst^＋ of NSCs from PO were58.81% ±9.69%（n=10）和11 22.85% ±4.36% （n = 10） while the ones for P2 were 27.84% ±5.70% （n = 10）and 49.32% ± 6.95% （ n = 10,P = 0. 000）. Conclusions By passaging the proliferation and apoptosis of NSCs decreases and they differentiate into less neurons and more astrocytes.
出处
《医学研究杂志》
2007年第1期62-65,共4页
Journal of Medical Research
关键词
神经干细胞
传代
增生
分化
凋亡
Neural stem cell
Passaging proliferation
Differentiation
Apoptosis
