Adenoviral-mediated gene transfer into ex vivo expanded human bone marrow mesenchymal progenitor cells
Adenoviral-mediated gene transfer into ex vivo expanded human bone marrow mesenchymal progenitor cells
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DOI：
10.1016/s0301-472x(00)00134-x
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发表时间：
2000-04-01
影响因子：
2.6
通讯作者：
Minguell, JJ
中科院分区：
文献类型：
--
作者：
Conget, PA;Minguell, JJ
Objective.人骨髓间充质祖细胞（mesenchymal progenitor cells，MPC）具有良好的分化特性和体外扩增的能力，被认为是将外源基因导入骨髓或其他间充质组织的理想靶细胞。在这项研究中，我们研究了用腺病毒载体（Adv），方法。将MPC离体扩增并在CMV启动子的控制下用含有复制缺陷型Adv-containing报告基因（lacZ或GFP）转导。流式细胞术检测Adv-粘附受体（CAR）和Adv-内化受体（integrin α v）的表达和参与情况。结果：转基因表达分析显示，只有19% ± 3%的细胞高水平表达转基因，MPC表达Adv感染所需的粘附受体和内化受体。虽然整联蛋白α v β 3和α v β 5由所有MPC表达，但CAR仅由一部分小尺寸细胞表达。针对CAR和α v β 5的抗体，但不针对α v β 3的抗体，阻断了Adv-mediated基因转移到MPC中，表明CAR和α v β 4是感染所必需的。因为与CAR相比，α v β 5在MPC中过表达，结果表明Adv-mediated基因转移到MPC中的效率取决于CAR表达水平。这些发现表明，Adv可用于工程化具有高水平转基因表达的离体扩增的人间充质祖细胞的亚群。(C)2000年国际实验线虫学会。出版社：Elsevier Science Inc.
Objective. Based on their differentiation properties and facilely of ex vivo expansion, human bone marrow mesenchymal progenitor cells (MPC), are considered as attractive targets to deliver foreign genes to the hone marrow or other mesenchymal tissues. In this study we investigated the feasibility of transduce MPC with adenoviral vectors (Adv),Methods. MPC were expanded ex vivo and transduced with replication-defective Adv-containing reporter genes (lacZ or GFP) under the control of CMV promoter. Transfection efficiency was assessed by microscopical scoring or by now cytometry, Expression and involvement of Adv-attachment (CAR) and Adv-internalization (integrins alpha v) receptors were evaluated by flow cytometric studies.Results, Transgene expression analysis showed that only 19% +/- 3% of cells expressed the transgenes at high levels, MPC express the attachment and internalization receptors required for Adv infection. While integrins alpha v beta 3 and alpha v beta 5 are expressed by all MPC, CAR is solely expressed by a fraction of low size cells. Antibodies against CAR and alpha v beta 5, but not against alpha v beta 3, blocked Adv-mediated gene transfer into MPC, showing that CAR and alpha v beta 4 are required for infection, Because alpha v beta 5, as compared with CAR, is overexpressed in MPC, the results suggest that the efficiency of Adv-mediated gene transfer into MPC depends on the level of CAR expression,Conclusion. These findings demonstrate that Adv may be useful to engineer a subpopulation of ex vivo expanded human mesenchymal progenitors, with a high level of transgene expression. (C) 2000 International Society for Experimental Nematology. Published by Elsevier Science Inc.