羧基荧光素乙酰乙酸与四甲基偶氮唑盐法检测人脐血间充质干细胞刺激脾淋巴细胞增殖作用的比较  

Compararative investigation of proliferation of lymphocytes stimulated by human umbilical cord blood mesenchymal stem cells with CFSE and MTT

作　　者：孙海梅[1] 季凤清[1] 王屹[2] 王丹妮[1] 赵春礼[3] 杨慧[2] 

机构地区：[1]首都医科大学组织学与胚胎学教研室 [2]中国康复研究中心检验科,北京100068 [3]首都医科大学北京神经科学研究所,北京100069

出　　处：《解剖学报》2009年第4期680-684,共5页Acta Anatomica Sinica

基　　金：国家重点基础研究发展计划(973);北京市教委科技发展计划(KM200610025005);北京市自然基金资助项目(7082017)

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...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 earboxyfluoreseein dideetate （CFSE） and MTY. Methods To prepare LCs from SD rat and divided into four group stimulating ceils： 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.

关 键 词：脐血间充质干细胞 淋巴细胞 增殖 羧基荧光素乙酰乙酸 四甲基偶唑盐比色法 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学]