【摘要】 目的比较羧基荧光素乙酰乙酸(CFSE)与四甲基偶氮唑盐(MTT)比色法检测人脐血间充质干细胞(CB-MSCs)和神经分化的CB-MSCs对大鼠脾淋巴细胞(LCs)刺激作用的敏感程度。方法分别制备刺激细胞和LCs,将刺激细胞分为4组:1.CB-MSCs组;2.Dif-CB-HSCs组:脐血间充质干细胞培养7d后维甲酸(RA)处理4d诱导其神经分化细胞;3.SH-SY5Y组:人源SH-SY5Y细胞(人神经母细胞瘤细胞系)作为阳性对照组;4.Auto-LC组:自体大鼠LCs作为阴性对照组。分别将上述各组刺激细胞与LCs进行混合培养,分别通过酶标仪(MTT法),流式细胞术(CFSE法)检测LCs增殖情况(n=3)。结果CFSE法和MTT法检测结果均显示,CB-MSCs组和Dif-CB-MSCs组刺激LCs增殖明显弱于阳性对照组(SH-SY5Y细胞组),但CFSE法检测结果显示,CB-MSCs组和Dif-CB-MSCs组细胞刺激LCs增殖百分率高于阴性对照Auto-LC组,MTT法检测结果显示,CB-MSCs组和Dif-CB-MSCs组的吸光度(A)值稍低于阴性对照Auto-LC组。结论CFSE法比MTT法能更加客观地反映淋巴细胞增殖情况,在实际应用中更具有优势。更多还原

【Abstract】 Objective To compare the sensitivities of the stimulation effect of human cord blood mesenchymal stem cells(CB-MSCs) and CB-MSCs of neuronal differentiation to lymphocytes(LCs) detected with carboxyfluorescein didcetate(CFSE) and MTT. Methods To prepare LCs from SD rat and divided into four group stimulating cells: 1.CB-MSCs;2.Dif-CB-HSCs;3.SH-SY5Y(positive control);4.Auto-LC(negative control).Stimulating cells were respectively Co-cultured with LCs. The proliferation of LCs was detected with MTT and CFSE(n=3). Results CB-MSCs and Dif-CB-HSCs stimulated LCs to proliferate more weakly than positive control detected with MTT and CFSE.The quantity of proliferation of lymphocytes Co-cultured with CB-MSCs and Dif-CB-HSCs were higher than that of Auto-LC detected with CFSE.But MTT OD value of CB-MSCs and Dif-CB-HSCs was a little lower than that of Auto-LC.Statistical analysis results showed no significant difference.Conclusion CFSE can reflect proliferation status of lymphocytes better than MTT.CFSE shows more advantages in practical use.更多还原