Cardiac applications of second harmonic generation (SHG) microscopy
Cardiac applications of second harmonic generation (SHG) microscopy
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DOI：
10.1117/12.2510387
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发表时间：
2019-02
期刊：
影响因子：
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通讯作者：
Che-Wei Chang;Hannah A. Ledford;Hillary K J Kao;Xiao-Dong Zhang;N. Chiamvimonvat;D. Lieu;J. Chan
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文献类型：
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作者：
Che-Wei Chang;Hannah A. Ledford;Hillary K J Kao;Xiao-Dong Zhang;N. Chiamvimonvat;D. Lieu;J. Chan
人诱导多能干细胞衍生的心肌细胞 (iPSC-CM) 是用于心脏应用的无限离体心脏细胞供应。纯 iPSC-CM 群体的建立对于人类疾病建模、患者特异性干细胞治疗、人体移植和药物开发等下游医学应用至关重要。然而，一个重大挑战是由于缺乏特定的 iPSC-CM 标记，缺乏既定的纯化方法来根据表型、成熟度和亚型分离 iPSC-CM 群体。去除可能形成畸胎瘤的多能干细胞、诱发心律失常的未成熟细胞和起搏细胞以及其他非 CM 的能力对于工程组织具有所需的细胞成分极其重要，这些细胞成分既可以安全地用于人体移植，又可以准确地复制心脏功能。现代纯化技术要么特异性低，要么需要基因改造。我们提出，已知源自心肌细胞中的肌节肌球蛋白丝的二次谐波产生 (SHG) 信号可以成为用于识别 iPSC-CM 的高度特异性、无标记标记。在这里，我们演示了使用 SHG 显微镜来表征 iPSC-CM 及其亚型。
Human induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) are an unlimited ex vivo supply of heart cells for cardiac applications. The establishment of pure iPSC-CMs populations is crucial for downstream medical applications such as human disease modeling, patient-specific stem cell therapy, human transplantation, and drug development. However, a significant challenge is the lack of an established purification method to isolate populations of iPSC-CMs by their phenotype, maturity, and subtype due to the lack of specific iPSC-CM markers. The ability to remove potentially teratoma forming pluripotent stem cells, arrhythmia inducing immature and pacemaking cells, and other non-CMs is extremely important for engineering tissues with desired cell compositions that are both safe for human transplantation and that can accurately replicate cardiac functions. Contemporary purification techniques have either low specificity or require genetic modification. We have proposed that second harmonic generation (SHG) signals, which are known to originate from the sarcomeric myosin filaments in cardiomyocytes, can be a highly specific, labelfree marker for identifying iPSC-CMs. Here, we demonstrate the use of SHG microscopy for characterizing iPSC-CMs and their subtypes.