首页 > 核心刊物 > 中国生物工程杂志 2017.6 Vol 37, No.6 > HIV-1 Tat蛋白抑制骨髓间充质干细胞的造血支持功能

HIV-1 Tat蛋白抑制骨髓间充质干细胞的造血支持功能

袁雅红, 赵珊珊, 王小莉, 腾智平, 李东升, 曾毅

1. 北京工业大学生命科学与生物工程学院 北京 100124 2. 湖北医药学院附属太和医院胚胎干细胞研究湖北省重点实验室 十堰 442000 3. 中国预防医学科学院病毒所 北京 100052

 
Key Words:  造血干细胞; 造血支持; HIV-1; 骨髓间充质干细胞; Tat蛋白
 

Abstract

目的:通过研究人类免疫缺陷病毒1型(HIV-1) Tat蛋白对骨髓间充质干细胞(BMSC)造血支持功能的影响,进一步揭示HIV-1感染者造血损伤的机理。方法:原代培养BMSC,流式检测其表面标志,诱导分化鉴定其多向分化潜能;免疫磁珠分选造血干细胞(HSC),流式检测其纯度;HIV-1 Tat蛋白添加到培养基中培养20天的BMSC(BMSCTat)与对照BMSC (BMSCCon)分别作为滋养层与HSC分6组进行共培养,随后计数各组造血细胞总数,诱导分化检测造血细胞集落形成能力;RT-PCR检测BMSCTat和BMSCCon造血相关因子mRNA的表达强度,ELISA检测BMSCTat和BMSCCon条件培养液中造血相关因子GM-CSF及IL-6的浓度。结果:经鉴定成功培养获得原代BMSC;免疫磁珠分选的HSC纯度可达95%以上;分6组共培养进行比较,以BMSCTat为滋养层培养的造血细胞总数及造血细胞形成的集落总数均明显少于以BMSCCon为滋养层;BMSCTat的造血相关因子的mRNA的表达明显弱于BMSCCon,BMSCTat的条件培养液中GM-CSF和IL-6的浓度均明显低于BMSCCon。结论:HIV-1 Tat蛋白对BMSC的造血支持功能有明显的抑制作用。


 

Abstract

Objective:In order to further reveal the mechanism of hematopoietic injury among people infected with HIV-1, The effect of HIV-1 Tat protein on hematopoiesis support function of bone marrow mesenchymal stem cells(BMSC) was researched. Methods:The bone marrow mesenchymal stem cells were primary cultured and their characters were identified by detecting differentiation potential and immune phenotype. Hematopoietic stem cells (HSC) were sorted with immunomagnetic beads and the purity were detected with flow cytometry. As the feeder layer, the control BMSC (BMSCCon) and the experiment BMSC which have been added HIV-1Tat protein into the medium for culturing 20 days (BMSCTat) were respectively co-cultured with HSC and divided into six groups. Then, a series of detections were carried out, the total numbers of hematopoietic cells were counted, hematopoietic cells clonogenic ability were assayed, hematopoietic growth fator mRNA expression of BMSCCon and BMSCTat were detected with RT-PCR, GM-CSF and IL-6 concentration in the condition medium of BMSCCon and BMSCTat were detected with ELISA. Results:Identification results show that the BMSC were successfully cultured.The purity of HSC sorted by immunomagnetic beads can reach more than 95%. The six groups were compared with each other and the results showed that both the total numbers of hematopoietic cells and the total numbers of hematopoietic cell colony in the groups when BMSCTat were as the feeder layer were significantly less than those in the groups when BMSCCon were as the feeder layer. The hematopoietic growth fator mRNA expression of BMSCTat were significantly weaker than that of BMSCCon. The GM-CSF and IL-6 concentration in the condition medium of BMSCTat were significantly lower than that of BMSCCon. Conclusions:The HIV1 Tat protein was confirmed to inhibit the hematopoiesis support function of bone marrow mesenchymal stem cells.

PDF全文


过刊检索
年:
月:
卷:
期刊号:
文章检索
标题:
作者:
作者单位:
关键字:
摘要:

期刊简介
 
主办单位:
中国生物工程学会(CSBT)、中国生物技术发展中心(CNCBD)、中国科学院文献情报中心。
编委会:
最新一届编辑委员会由国家生物技术研究开发计划的评审专家及有关院士组成,代表中国生物技术的最高学术水平。
编委会主任:陈志南
通信地址:北京市中关村北四环西路33号《中国生物工程杂志》编辑部(100190)
国内刊号:CN11-4816/Q
国际刊号:ISSN1671-8135
邮发代号:82-673