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首页> 外文期刊>Stem cells and development >Do Induced Pluripotent Stem Cell Characteristics Correlate with Efficient In Vitro Smooth Muscle Cell Differentiation? A Comparison of Three Patient-Derived Induced Pluripotent Stem Cell Lines
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Do Induced Pluripotent Stem Cell Characteristics Correlate with Efficient In Vitro Smooth Muscle Cell Differentiation? A Comparison of Three Patient-Derived Induced Pluripotent Stem Cell Lines

机译：是否诱导多能干细胞特征与有效的体外平滑肌细胞分化相关？ 三种患者衍生诱导多能干细胞系的比较

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Human induced pluripotent stem cells (iPSCs) have the potential to repair/regenerate smooth muscle cells (SMCs) in different organs. However, there are many challenges in their translation to clinical therapies. In this study, we describe our observations of in vitro SMC differentiation in three iPSC lines derived from human fibroblasts using retroviral, episomal, and mRNA/miRNA reprogramming methods. We sought to elucidate correlations between differentiation characteristics and efficiencies that can facilitate large-scale production of differentiated cells for clinical applications, and to report differences in pluripotency marker expression in differentiated cells from different iPSC lines. A standardized SMC differentiation protocol was used to induce the CD31(+)/CD34(+) vascular progenitor cell phenotype. These were sorted by magnetic-activated (MACS) and fluorescence-activated cell sorting (FACS), and then treated with PDGF-BB and smooth muscle growth medium for further differentiation into smooth muscle progenitor cells (pSMCs). The expression of SMC and pluripotency markers in early- and late-passage (P1 and P4) pSMCs was analyzed. A total of 36 differentiation runs was performed on the three patient iPSC lines. All pSMC populations expressed SMC markers and Ki67 consistent with the progenitor phenotype. Initial iPSC density correlated positively with the sorted cell FACS efficiency, and this correlation could be fit to a quadratic equation. We also observed that a specific "honey-comb" pattern of the starting cultured iPSCs cultured correlated with higher efficiency in all three iPSC lines. Pluripotency marker expression decreased significantly to nearly undetectable levels in all three lines. There was no significant change in SMC and pluripotent marker expression between passage 1 and 4. In summary, our observations suggest that the method of iPSC reprogramming does not affect iPSC differentiation into pSMCs. Protocol efficiency can be modeled mathematically and coupled with the initial honeycomb cell pattern to optimize production of large cell numbers for clinical therapies.
机译：人诱导多能干细胞（IPSC）具有潜力在不同器官中修复/再生平滑肌细胞（SMC）。然而，他们与临床疗法的翻译有许多挑战。在这项研究中，我们描述了使用逆转录病毒，卵象和mRNA / miRNA重编程方法衍生自人成纤维细胞的三种IPSC线体外SMC分化的观察。我们试图阐明分化特征和效率之间可以促进临床应用的大规模产生的效率之间的相关性，并在不同IPSC线中报告不同IPSC线的分化细胞中多能性标志物表达的差异。标准化的SMC分化方案用于诱导CD31（+）/ CD34（+）血管祖细胞表型。这些由磁性活化（MACs）和荧光激活的细胞分选（FACS）分类，然后用PDGF-BB和平滑肌生长培养基处理，以进一步分化为平滑肌祖细胞（PSMC）。分析了SMC和多能性标志物在早期和晚期通道（P1和P4）PSMC中的表达。在三个患者IPSC线上进行了总共36个差异运行。所有PSMC群体表达了SMC标记和KI67与祖细胞表型一致。初始IPSC密度随着分类的小区FACS效率而正相关，并且这种相关性可以适合二次方程。我们还观察到，在所有三个IPSC线路中，开始培养的IPSCS的特定“蜂蜜梳”模式与较高效率相关。多能性标志物表达在所有三条线中显着降低到几乎不可检测的水平。在第1和4段和4之间没有显着变化和多能标记表达。总之，我们的观察结果表明，IPSC重新编程的方法不会影响IPSC分化为PSMC。协议效率可以在数学上进行建模并与初始蜂窝单元格模式耦合，以优化临床疗法的大细胞数的产生。

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