Shh协同骨髓间充质干细胞对造血干细胞增殖的影响  

Effect of Shh and BM-MSC Synergism on the Proliferation of Hematopoietic Stem Cells

作　　者：郭静 汪姝玥 朱小凤 李书坛 林凡莉 李晓明 黄纯兰 GUO Jing;WANG Shu-Yue;ZHU Xiao-Feng;LI Shu-Tan;LIN Fan-li;LI Xiao-Ming;HUANG Chun-Lan(Department of Hematology,Affiliated Hospital of Southwest Medical University,Luzhou 646000,Sichuan Province,China)

机构地区：[1]西南医科大学附属医院血液内科,四川泸州646000

出　　处：《中国实验血液学杂志》2018年第5期1523-1530,共8页Journal of Experimental Hematology

基　　金：国家自然基金课题(81450030);四川省2015年度留学回国人员科技活动择优资助人选重点项目资助;四川省教育厅重大培育项目(14CZ0017)

摘　　要：目的:探讨Shh协同骨髓间充质干细胞在体外非接触共培养中对造血干细胞增殖的影响及其机制。方法:采用全骨髓贴壁培养法体外培养间充质干细胞,采用miniMACS磁珠分选仪分选人骨髓CD34+细胞,流式细胞术鉴定两种细胞纯度;将CD34+细胞与MSC分别种至Transwell上下室进行非接触共培养,并添加外源性shh蛋白进行干预。收集非接触共培养d 7的样本检测HSC及MSC细胞数量、RNA总量、HSC的ki67的mRNA表达量,以了解HSC增殖情况;测量HSC细胞Tie-2 mRNA表达量和MSC的VEGF、Ang-1 mRNA表达量,以了解细胞因子的表达情况。结果:非接触共培养d 7,共培养组2种细胞数量、RNA总量以及ki67、Tie-2、VEGF、Ang-1相对表达量增加并呈以下趋势:MSC+HSC组和shh+HSC组均高于HSC组(P <0.05),MSC+shh+HSC组高于MSC+HSC组(P <0.05)。结论:shh可协同MSC体外非接触共培养促进HSC增殖,其机制可能与血管生成因子相关。Objective:To study the effect and mechanism of shh and mesenchymal stem cell(MSC)synergism on the proliferation of hematopoietic stem cells in noninvasive co-culture system in vitro.Methods:The mesenchymal stem cells were cultured in vitro,CD34+cells were sorted by mini MACS magnetic bead separator,flow cytometry was used to identify the purity of 2 cells.CD34+cells and MSCs were seeded to upper and low of transwell respecibely for noncontact coculture,and add exogenous shh protein for intervenece.The number of MSCs and HSCs,the total amount of RNA,the expression of ki67 and Tie-2 mRNA of HSC,the expression of VEGF and Ang-1 mRNA of MSC were detected for investigating the condition of cell proliferation and the expression of angiogenic factors.Results:The total number of cells,the total amount of RNA and the relative expression of ki67,Tie-2,VEGF and Ang-1 in non-contact co-culture group increased and showed the following trends on the 7th day:the above-mentioned indexes in group MSC+HSC,group shh+HSC were higher than those in group HSC,while those in MSC+shh+HSC Group was higher than those in MSC+HSC and shh+HSC group.Conclusion:Angiogenic factors help MSC to proliferate HSC and amplify the CD34+hematopoietic stem/progenitor cells by shh and MSC synergism in vitro coculture system which may be related with angiogenic factors.

关 键 词：造血干细胞 间充质干细胞 Sonic HEDGEHOG 血管生成因子 

分 类 号：R331.2[医药卫生—人体生理学]