The Francis Crick Institute, London, UK.
NHLI, Imperial College London, London, UK.
Cell Rep Methods. 2023 Apr 24;3(4):100456. doi: 10.1016/j.crmeth.2023.100456.
Decreased left ventricle (LV) function caused by genetic mutations or injury often leads to debilitating and fatal cardiovascular disease. LV cardiomyocytes are, therefore, a potentially valuable therapeutical target. Human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) are neither homogeneous nor functionally mature, which reduces their utility. Here, we exploit cardiac development knowledge to instruct differentiation of hPSCs specifically toward LV cardiomyocytes. Correct mesoderm patterning and retinoic acid pathway blocking are essential to generate near-homogenous LV-specific hPSC-CMs (hPSC-LV-CMs). These cells transit via first heart field progenitors and display typical ventricular action potentials. Importantly, hPSC-LV-CMs exhibit increased metabolism, reduced proliferation, and improved cytoarchitecture and functional maturity compared with age-matched cardiomyocytes generated using the standard WNT-ON/WNT-OFF protocol. Similarly, engineered heart tissues made from hPSC-LV-CMs are better organized, produce higher force, and beat more slowly but can be paced to physiological levels. Together, we show that functionally matured hPSC-LV-CMs can be obtained rapidly without exposure to current maturation regimes.
由于基因突变或损伤导致的左心室（LV）功能下降常导致使人衰弱和致命的心血管疾病。因此，LV 心肌细胞是一种有潜在价值的治疗靶点。人多能干细胞衍生的心肌细胞（hPSC-CMs）既不均匀也不成熟，这降低了它们的效用。在这里，我们利用心脏发育知识来专门指导 hPSC 向 LV 心肌细胞分化。正确的中胚层模式形成和视黄酸途径阻断对于产生近同质的 LV 特异性 hPSC-CMs（hPSC-LV-CMs）至关重要。这些细胞通过第一心区祖细胞过渡，并显示出典型的心室动作电位。重要的是，与使用标准 WNT-ON/WNT-OFF 方案生成的年龄匹配的心肌细胞相比，hPSC-LV-CMs 表现出更高的代谢率、更低的增殖率以及更好的细胞结构和功能成熟度。同样，由 hPSC-LV-CMs 制成的工程化心脏组织组织更加有序，产生的力更高，跳动速度更慢，但可以起搏到生理水平。总之，我们表明可以在不暴露于现有成熟方案的情况下快速获得功能成熟的 hPSC-LV-CMs。
