过氧化氢酶对人造血干细胞在免疫缺陷小鼠中植入的作用  

Effects of Catalase on the Engraftment of Human Hematopoietic Stem Cells in NOD/SCID Mice

作　　者：师鸣 胡林萍 张孝兵 袁卫平 程涛 

机构地区：[1]中国医学科学院、北京协和医学院血液学研究所、血液病医院,实验血液学国家重点实验室,天津300020

出　　处：《中国实验血液学杂志》2015年第1期183-189,共7页Journal of Experimental Hematology

基　　金：天津市科学技术委员会自然基金(11JCZDJZ27900,13JCYBJC39400);国家自然科学基金(81400150,81328004,81330015)

摘　　要：目的:研究人脐带组织来源的间充质干细胞(umbilical cord-derived mesenchymal stem cells,UC-MSC)过表达过氧化氢酶(catalase,CAT)基因对人造血干细胞(hematopoietic stem cells,HSC)移植作用的影响。方法:用携带GFP空载体与GFP-CAT载体的逆转录病毒感染UC-MSC,流式细胞术分选得到MSC-GFP和MSC-GFP-CAT两种细胞株,检测两种细胞株中CAT的mRNA表达水平和过氧化氢酶的酶活力;将其与人脐血CD34+细胞体外共培养;与人Lin-CD34+CD38-CD45RA-CD90+CD49f+Rholow细胞共同移植到NOD/SCID小鼠胫骨中,移植12周后,检测骨髓中的人CD45+细胞比例。结果:感染48 h后能够观察到UC-MSC发出绿色荧光,并能够稳定传代;流式细胞术分选后,MSC-GFP的纯度达到97.6%,MSC-GFP-CAT达到96.8%;MSC-GFP-CAT细胞中CAT的mRNA水平为对照组的23.9倍;体外检测UC-MSC、MSC-GFP和MSC-GFP-CAT细胞的酶活力分别为19.5、20.3、74.1 Unit;在共培养实验中,各组中的CD34+细胞比例无显著差异;在共移植实验中,HSC、MSC-GFP和MSC-GFP-CAT组中人CD45+细胞的比例分别为(3.22±3.1)%、(4.26±3.56)%和(7.37±4.51)%,MSC-GFP-CAT组的植入率为对照组MSC-GFP的1.7倍。结论:MSC-GFP-CAT对人HSC体外扩增无影响;MSC-GFP-CAT与人HSC共移植能够提高人HSC的植入率。Objective ： To investigate the effect of catalase （CAT） on engraftment of human hematopoietic stem cells （HSC） by co-transplanting umbilical cord-derived mesenchymal stem cells （UC-MSC） with over-expressed CAT and human HSC into NOD/SCID mice. Methods： The UC-MSC cultured in vitro were transfected by the retrovirus containing green fluorescent protein （GFP） and GFP-CAT genes respectively. MSC-GFP and MSC-GFP-CAT cell lines were sorted by flow cytometry. Co-culture and co-transplant experiments were performed to detect the effects of CAT on expansion and engraftment of human HSC. Results： The percentage of GFP ＋ cells were approximately 97.6% and 96.8% after sorting. The mRNA expression of CAT in MSC-GFP-CAT was 23.9-fold higher than that in UC-MSC. The activity of CAT in UC-MSC, MSC-GFP, MSC-GFP-CAT cells were 19.5, 20.3 and 74.1 Unit respectively. There was no significant differences in the percentage of CD34 ~ cells between 3 groups in co-culture experiment. And the percentage of human CD45＋ ceils in NOD/SCID mice were （ 3.22 ± 3.1 ） % , （4.26 ± 3.56 ） % and （ 7.37 ± 4.51 ） % respectively. Conclusion： MSC-GFP-CAT significantly improves the engraftment of human HSC in NOD/SCID mice, whereas coculture with the MSC-GFP-CAT can not promote the expansion of HSC in vitro.

关 键 词：造血干细胞 间充质干细胞 过氧化氢酶 植入率 NOD/SCID小鼠 

分 类 号：R457.7[医药卫生—治疗学]