An intrinsic, label-free signal for identifying stem cell-derived cardiomyocyte subtype
An intrinsic, label-free signal for identifying stem cell-derived cardiomyocyte subtype
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DOI：
10.1002/stem.3127
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发表时间：
2020-03-01
期刊：
影响因子：
5.2
通讯作者：
Chan, James W.
中科院分区：
文献类型：
--
作者：
Chang, Che-Wei;Kao, Hillary K. J.;Chan, James W.
人诱导多能干细胞（hiPSC）衍生的心肌细胞具有许多有前景的应用，包括损伤心肌的再生、心血管疾病建模和药物心脏毒性筛选。目前的分化方案产生了一个异质细胞群，包括多能干细胞和不同的心脏亚型（起搏和收缩细胞）。纯化这些细胞并获得明确定义的、受控的细胞组成的能力对于许多下游应用是重要的;然而，目前还没有建立和可靠的方法来鉴定hiPSC衍生的心肌细胞及其亚型。在这里，我们证明了直接从肌球蛋白杆束产生的二次谐波产生（SHG）信号可以是用于识别hiPSC衍生的心肌细胞的无标记的内在光学标记。观察到SHG信号强度与心脏亚型之间的直接相关性，起搏器样细胞通常表现出比心房和心室样心肌细胞低约70%的信号强度。这些发现表明，起搏器样细胞可以从异质性人口中分离，通过选择一个SHG强度阈值标准。该工作为开发用于纯化hiPSC衍生的心肌细胞及其亚型的基于SHG的高通量流式分选仪奠定了基础。
Human-induced pluripotent stem cell (hiPSC)-derived cardiomyocytes have many promising applications, including the regeneration of injured heart muscles, cardiovascular disease modeling, and drug cardiotoxicity screening. Current differentiation protocols yield a heterogeneous cell population that includes pluripotent stem cells and different cardiac subtypes (pacemaking and contractile cells). The ability to purify these cells and obtain well-defined, controlled cell compositions is important for many downstream applications; however, there is currently no established and reliable method to identify hiPSC-derived cardiomyocytes and their subtypes. Here, we demonstrate that second harmonic generation (SHG) signals generated directly from the myosin rod bundles can be a label-free, intrinsic optical marker for identifying hiPSC-derived cardiomyocytes. A direct correlation between SHG signal intensity and cardiac subtype is observed, with pacemaker-like cells typically exhibiting ~70% less signal strength than atrial- and ventricular-like cardiomyocytes. These findings suggest that pacemaker-like cells can be separated from the heterogeneous population by choosing an SHG intensity threshold criteria. This work lays the foundation for developing an SHG-based high-throughput flow sorter for purifying hiPSC-derived cardiomyocytes and their subtypes.