Use of Adeno-Associated Virus to Enrich Cardiomyocytes Derived from Human Stem Cells
Use of Adeno-Associated Virus to Enrich Cardiomyocytes Derived from Human Stem Cells
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DOI：
10.1089/humc.2015.052
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发表时间：
2015-09-01
影响因子：
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通讯作者：
Childers, Martin K.
中科院分区：
文献类型：
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作者：
Guan, Xuan;Wang, Zejing;Childers, Martin K.
来源于人诱导多能干细胞（iPSC）的心肌细胞显示出作为自体供体细胞治疗心脏病的巨大前景。这种方法的主要技术障碍是可用的诱导方法通常产生具有低百分比心肌细胞的异质细胞群。在这里，我们描述了一个心脏富集的方法，使用非整合腺相关病毒（AAV）。我们首先检查了几种AAV血清型选择性地扩增iPSC衍生的心肌细胞的能力。结果显示，在广泛使用的7种血清型中，AAV1表现出最高的体外转导效率。接下来，用表达新霉素抗性基因的药物选择性AAV 1转导分化的iPSC衍生物。用G418进行的选择在2周内使心肌细胞分数从27%富集至57%。与其他富集策略如整合遗传选择、线粒体标记或表面标记细胞分选相比，本文所述的这种简单的AAV方法绕过了抗体或染料标记。这些发现提供了通过利用AAV的内在组织向性进行大规模心肌细胞富集的概念证据。
Cardiomyocytes derived from human induced pluripotent stem cells (iPSCs) show great promise as autologous donor cells to treat heart disease. A major technical obstacle to this approach is that available induction methods often produce heterogeneous cell population with low percentage of cardiomyocytes. Here we describe a cardiac enrichment approach using nonintegrating adeno-associated virus (AAV). We first examined several AAV serotypes for their ability to selectively transduce iPSC-derived cardiomyocytes. Results showed that AAV1 demonstrated the highest in vitro transduction efficiency among seven widely used serotypes. Next, differentiated iPSC derivatives were transduced with drug-selectable AAV1 expressing neomycin resistance gene. Selection with G418 enriched the cardiac cell fraction from 27% to 57% in 2 weeks. Compared with other enrichment strategies such as integrative genetic selection, mitochondria labeling, or surface marker cell sorting, this simple AAV method described herein bypasses antibody or dye labeling. These findings provide proof of concept for large-scale cardiomyocyte enrichment by exploiting AAV's intrinsic tissue tropism.