Journal of Molecular and Cellular Cardiology ( IF 4.9 ) Pub Date : 2021-08-13 , DOI: 10.1016/j.yjmcc.2021.08.005 Lili Wang 1 , Yuko Wada 1 , Nimer Ballan 2 , Jeffrey Schmeckpeper 1 , Jijun Huang 3 , Christoph Daniel Rau 3 , Yibin Wang 3 , Lior Gepstein 4 , Bjorn C Knollmann 1
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Background
Human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) have emerged as a promising tool for disease modeling and drug development. However, hiPSC-CMs remain functionally immature, which hinders their utility as a model of human cardiomyocytes.
Objective
To improve the electrophysiological maturation of hiPSC-CMs.
Methods and results
On day 16 of cardiac differentiation, hiPSC-CMs were treated with 100 nmol/L triiodothyronine (T3) and 1 μmol/L Dexamethasone (Dex) or vehicle for 14 days. On day 30, vehicle- and T3 + Dex-treated hiPSC-CMs were dissociated and replated either as cell sheets or single cells. Optical mapping and patch-clamp technique were used to examine the electrophysiological properties of vehicle- and T3 + Dex-treated hiPSC-CMs. Compared to vehicle, T3 + Dex-treated hiPSC-CMs had a slower spontaneous beating rate, more hyperpolarized resting membrane potential, faster maximal upstroke velocity, and shorter action potential duration. Changes in spontaneous activity and action potential were mediated by decreased hyperpolarization-activated current (If) and increased inward rectifier potassium currents (IK1), sodium currents (INa), and the rapidly and slowly activating delayed rectifier potassium currents (IKr and IKs, respectively). Furthermore, T3 + Dex-treated hiPSC-CM cell sheets (hiPSC-CCSs) exhibited a faster conduction velocity and shorter action potential duration than the vehicle. Inhibition of IK1 by 100 μM BaCl2 significantly slowed conduction velocity and prolonged action potential duration in T3 + Dex-treated hiPSC-CCSs but had no effect in the vehicle group, demonstrating the importance of IK1 for conduction velocity and action potential duration.
Conclusion
T3 + Dex treatment is an effective approach to rapidly enhance electrophysiological maturation of hiPSC-CMs.
中文翻译：
三碘甲状腺原氨酸和地塞米松改变钾通道表达并促进人诱导多能干细胞衍生心肌细胞的电生理成熟
背景
人诱导的多能干细胞衍生心肌细胞 (hiPSC-CM) 已成为疾病建模和药物开发的有前途的工具。然而，hiPSC-CM 在功能上仍不成熟，这阻碍了它们作为人类心肌细胞模型的实用性。
客观的
提高 hiPSC-CM 的电生理成熟度。
方法和结果
在心脏分化的第 16 天，hiPSC-CM 用 100 nmol/L 三碘甲腺原氨酸 (T3) 和 1 μmol/L 地塞米松 (Dex) 或载体处理 14 天。在第 30 天，载体和 T3 + Dex 处理的 hiPSC-CM 被解离并重新接种为细胞片或单细胞。光学映射和膜片钳技术用于检查载体和 T3 + Dex 处理的 hiPSC-CM 的电生理特性。与载体相比，T3 + Dex 处理的 hiPSC-CM 具有更慢的自发搏动率、更超极化的静息膜电位、更快的最大上冲速度和更短的动作电位持续时间。自发活动和动作电位的变化是由降低的超极化激活电流 (I f ) 和增加的内向整流钾电流 (I K1)、钠电流 (I Na )，以及快速和缓慢激活的延迟整流钾电流（分别为 I Kr和 I Ks）。此外，T3 + Dex 处理的 hiPSC-CM 细胞片 (hiPSC-CCS) 表现出比载体更快的传导速度和更短的动作电位持续时间。在 T3 + Dex 处理的 hiPSC-CCS 中，100 μM BaCl 2对 I K1的抑制显着减慢了传导速度并延长了动作电位持续时间，但在载体组中没有影响，证明了 I K1对于传导速度和动作电位持续时间的重要性。
结论
T3 + Dex 处理是快速增强 hiPSC-CM 电生理成熟的有效方法。
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