Generation of highly purified human cardiomyocytes from peripheral blood mononuclear cell-derived induced pluripotent stem cells.
Generation of highly purified human cardiomyocytes from peripheral blood mononuclear cell-derived induced pluripotent stem cells.
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外周血单核细胞衍生的多能干细胞的高度纯化的人类心肌细胞产生。
DOI：
10.1371/journal.pone.0126596
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Hengstenberg C
中科院分区：
文献类型：
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作者：
Fuerstenau-Sharp M;Zimmermann ME;Stark K;Jentsch N;Klingenstein M;Drzymalski M;Wagner S;Maier LS;Hehr U;Baessler A;Fischer M;Hengstenberg C
诱导多能干细胞（iPS）在生理学研究中具有巨大的潜力。开发了一种新的方案，其将从外周血衍生iPS与优化的定向分化为心肌细胞和随后的代谢选择相结合。将人iPS细胞从活化的T细胞逆转录病毒去分化。随后优化的定向分化方案在分化的第16天产生30-45%的心肌细胞。衍生的心肌细胞表达适当的结构标志物，如心肌肌钙蛋白T，α-辅肌动蛋白和肌球蛋白轻链2（MLC 2 V）。在随后的乳酸代谢选择中，心肌细胞含量可以增加到90%以上。在代谢选择过程中心肌细胞的损失小于50%，而替代的基于表面抗体的选择程序导致高达80%的心肌细胞的损失。电生理学表征证实了典型的心脏特征以及衍生的心肌细胞群体内心室、心房和类心房动作电位的存在。我们的组合和优化方案非常稳健，适用于可扩展的心脏分化。它提供了一种简单且具有成本效益的方法，无需昂贵的设备即可产生大量高度纯化的功能性心肌细胞。它将进一步增强iPS细胞衍生的心肌细胞用于疾病建模、药物发现和再生医学的适用性。
Induced pluripotent stem (iPS) cells have an enormous potential for physiological studies. A novel protocol was developed combining the derivation of iPS from peripheral blood with an optimized directed differentiation to cardiomyocytes and a subsequent metabolic selection. The human iPS cells were retrovirally dedifferentiated from activated T cells. The subsequent optimized directed differentiation protocol yielded 30-45% cardiomyocytes at day 16 of differentiation. The derived cardiomyocytes expressed appropriate structural markers like cardiac troponin T, α-actinin and myosin light chain 2 (MLC2V). In a subsequent metabolic selection with lactate, the cardiomyocytes content could be increased to more than 90%. Loss of cardiomyocytes during metabolic selection were less than 50%, whereas alternative surface antibody-based selection procedures resulted in loss of up to 80% of cardiomyocytes. Electrophysiological characterization confirmed the typical cardiac features and the presence of ventricular, atrial and nodal-like action potentials within the derived cardiomyocyte population. Our combined and optimized protocol is highly robust and applicable for scalable cardiac differentiation. It provides a simple and cost-efficient method without expensive equipment for generating large numbers of highly purified, functional cardiomyocytes. It will further enhance the applicability of iPS cell-derived cardiomyocytes for disease modeling, drug discovery, and regenerative medicine.
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影响因子：
64.8
作者：
Itzhaki, Ilanit;Maizels, Leonid;Gepstein, Lior
通讯作者：
Gepstein, Lior
影响因子：
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作者：
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Maier, Lars S.
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通讯作者：
Daley GQ
影响因子：
64.8
作者：
通讯作者：
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