Adenovirus-mediated expression of the HO-1 protein within MSCs decreased cytotoxicity and inhibited apoptosis induced by oxidative stresses
Adenovirus-mediated expression of the HO-1 protein within MSCs decreased cytotoxicity and inhibited apoptosis induced by oxidative stresses
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DOI：
10.1007/s12192-011-0298-y
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发表时间：
2012-03-01
影响因子：
3.8
通讯作者：
Roudkenar, Mehryar Habibi
中科院分区：
文献类型：
--
作者：
Hamedi-Asl, Pejman;Halabian, Raheleh;Roudkenar, Mehryar Habibi
间充质干细胞（MSCs）在靶组织中存活和移植的能力可能导致有希望的治疗效果。然而，大多数MSC在移植后的最初几天内死亡的事实使细胞治疗复杂化。因此，有必要加强干细胞以承受微环境的严酷，以提高细胞治疗的功效。在这项研究中，我们操纵骨髓间充质干细胞表达细胞保护因子，血红素加氧酶-1（HO-1），以解决这个问题。通过TOPO克隆反应，将人HO-1全长cDNA克隆到TOPO载体中。然后，通过LR重组反应将构建体连接到网关适应的腺病毒表达载体。然后，在合适的哺乳动物细胞系中产生表达HO-1的重组病毒，并用于感染MSC。将HO-1工程化的MSC暴露于缺氧和氧化应激条件下，然后评价细胞的活力和凋亡。在MSC中检测到HO-1的瞬时表达。观察到HO-1表达可以保护MSC免受由缺氧和氧化应激条件引发的细胞死亡和凋亡。MSCs-HO-1保留了其分化成脂肪形成、软骨形成或成骨谱系的能力。这些发现可以作为一种策略，用于预防移植细胞死亡的MSC为基础的细胞治疗，是一个很好的示范细胞应激反应的理解可以用于实际应用。
The capacity of mesenchymal stem cells (MSCs) to survive and engraft in the target tissue may lead to promising therapeutic effects. However, the fact that the majority of MSCs die during the first few days following transplantation complicates cell therapy. Hence, it is necessary to strengthen the stem cells to withstand the rigors of the microenvironment to improve the efficacy of cell therapy. In this study, we manipulated MSCs to express a cytoprotective factor, heme oxygenase-1 (HO-1), to address this issue. Full-length cDNA of human HO-1 was isolated and cloned into TOPO vector by TOPO cloning reaction. Then, the construct was ligated to gateway adapted adenovirus expression vector by LR recombination reaction. Afterwards, the recombinant virus expressing HO-1 was produced in appropriate mammalian cell line and used to infect MSCs. The HO-1 engineered MSCs were exposed to hypoxic and oxidative stress conditions followed by evaluation of the cells' viability and apoptosis. Transient expression of HO-1 was detected within MSCs. It was observed that HO-1 expression could protect MSCs against cell death and the apoptosis triggered by hypoxic and oxidative stress conditions. The MSCs-HO-1 retained their ability to differentiate into adipogenic, chondrogenic, or osteogenic lineages. These findings could be applied as a strategy for prevention of graft cell death in MSCs-based cell therapy and is a good demonstration of how an understanding of cellular stress responses can be used for practical applications.