SDF-1/RUNX1融合蛋白介导的间充质干细胞对造血干细胞增殖和趋化作用的研究  

MSCs mediated by SDF-1/RUNX1 fusion protein induce the proliferation and chemotaxis of HSCs

作　　者：罗红春[1] 张红宾[2] 秦波[1] 汪嘉莉[1] 刘倩 

机构地区：[1]重庆医科大学附属第一医院感染科,400016 [2]重庆医科大学附属第一医院血液科,400016 [3]重庆市渝北区人民医院消化内科,401120

出　　处：《重庆医学》2012年第30期3129-3131,F0002,共4页Chongqing medicine

基　　金：重庆市卫生局医学科研计划基金资助项目(2010-2-095)

摘　　要：目的研究基质细胞衍生因子-1(SDF-1)/RUNX1融合蛋白介导的间充质干细胞(MSCs)对造血干细胞(HSCs)增殖和趋化能力的作用。方法分离培养人脐带源性MSCs和人脐血源性CD34+干细胞。重组腺病毒pAD-SDF-1-(GlySer)3-RUNX1-IRES-GFP感染MSCs,免疫荧光显微镜下观察感染效率,ELISA方法检测上清液中SDF-1和RUNX1的表达。流式细胞仪检测感染了重组腺病毒的MSCs对CD34+干细胞的扩增倍数,Transwell试验测定CD34+干细胞的迁移指数。结果重组腺病毒感染人脐带源性MSCs后,第3天的转染效率和SDF-1/RUNX1的表达最高,第7天也有较高表达,两者比较差异无统计学意义(P>0.05)。感染重组腺病毒的MSCs和CD34+干细胞共培养第3、7、14天,B1、C1组较A1组CD34+干细胞显著扩增(P<0.05)。培养第7、14天,B1组中CD34+干细胞的扩增倍数显著高于C1组(P<0.05)。Transwell试验中,B2、C2组的迁移指数明显高于A2组(P<0.05),并且C2组的迁移指数明显高于B2组,差异有统计学意义(P<0.05)。结论 SDF-1/RUNX1融合蛋白介导的MSCs可促进CD34+干细胞的增殖和归巢。Objective To investigate the proliferation and chemotaxis of HSCs influenced by MSCs which mediated by SDF-1/RUNX1 fusion protein.Methods We isolated and cultured MSCs derived from human umbilical cord and HSCs derived from human cord blood.The recombinated adenovirus pAD-SDF-1-（GlySer）3-RUNX1-IRES-GFP infected MSCs.The efficiency of infection was detected by fluorescence microscope.ELISA was used to assay the expression of SDF-1/RUNX1 in supernatant fluid.Then,we detected the proliferation and the immigration index of CD34^＋ stem cell influenced by MSCs infected by the recombinated adenovirus.Results After 3 days MSCs infected by recombinated adenovirus,the efficiency of transfection and the expression of SDF-1/RUNX1 were the highest.And after 7 days infection,they were still very high（P〈0.05）.There was no statistics difference.MSCs infected by recombinated adenovirus and CD34^＋ stem cell co-cultured,after 3,7,14 days,the proliferation of CD34^＋ stem cell in group B1,C1 were more significant than in group A1（P〈0.05）.After 7,14 days,the proliferation of CD34^＋ stem cell in group B1 was higher than in group C1（P〈0.05）.In transwell experience,the immigration indexs of CD34^＋ stem cell in group B2,C2 were higher than in group A2（P〈0.05）.Meanwhile,the immigration index of CD34^＋ stem cell in group C2 was obviously increased than in group B2（P〈0.05）.Conclusion MSCs mediated by SDF-1/RUNX1 fusion protein promote the proliferation and homing of CD34^＋ stem cell.

关 键 词：趋化因子CXCL12 RUNX1基因 间质干细胞 造血干细胞 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]