Optical Investigation of Action Potential and Calcium Handling Maturation of hiPSC-Cardiomyocytes on Biomimetic Substrates

International Journal of Molecular Sciences

Raffaele Coppini¹  Valentina Balducci¹  Laura Sartiani¹  Chiara Palandri¹  Elisabetta Cerbai¹  Lorenzo Santini¹  Michael Regnier²  Patrizia Benzoni³  Sara Landi³  Andrea Barbuti³  Cecilia Ferrantini⁴  Silvia Querceto⁴  Marianna Langione⁴  Chiara Tesi⁴  JosèManuel Pioner⁴  Corrado Poggesi⁴  DavidL. Mack⁵  Camilla Parmeggiani⁶  Flavia Lupi⁶  Bruno Grandinetti⁶  Daniele Martella⁶  Federico Ferrarese Lupi⁷  Luca Boarino⁷ 

[1] Department NeuroFarBa, University of Florence, 50134 Florence, Italy;Department of Bioengineering, University of Washington, Seattle, WA 98108, USA;Department of Biosciences, Università degli studi di Milano, 20137 Milan, Italy;Department of Experimental and Clinical Medicine, Division of Physiology, Università degli studi di Firenze, 50134 Florence, Italy;Department of Rehabilitation Medicine, University of Washington, Seattle, WA 98108, USA;European Laboratory for Non-Linear Spectroscopy (LENS), 50019 Florence, Italy;Istituto Nazionale di Ricerca Metrologica INRiM, 10129 Turin, Italy;

关键词: human induced pluripotent stem cells;    cardiomyocytes;    fluorescence;    maturation;    action potential;    calcium handling;    hydrogels;    long-term culture;   

DOI  :  10.3390/ijms20153799

来源: DOAJ

【 摘 要 】

Cardiomyocytes from human induced pluripotent stem cells (hiPSC-CMs) are the most promising human source with preserved genetic background of healthy individuals or patients. This study aimed to establish a systematic procedure for exploring development of hiPSC-CM functional output to predict genetic cardiomyopathy outcomes and identify molecular targets for therapy. Biomimetic substrates with microtopography and physiological stiffness can overcome the immaturity of hiPSC-CM function. We have developed a custom-made apparatus for simultaneous optical measurements of hiPSC-CM action potential and calcium transients to correlate these parameters at specific time points (day 60, 75 and 90 post differentiation) and under inotropic interventions. In later-stages, single hiPSC-CMs revealed prolonged action potential duration, increased calcium transient amplitude and shorter duration that closely resembled those of human adult cardiomyocytes from fresh ventricular tissue of patients. Thus, the major contribution of sarcoplasmic reticulum and positive inotropic response to β-adrenergic stimulation are time-dependent events underlying excitation contraction coupling (ECC) maturation of hiPSC-CM; biomimetic substrates can promote calcium-handling regulation towards adult-like kinetics. Simultaneous optical recordings of long-term cultured hiPSC-CMs on biomimetic substrates favor high-throughput electrophysiological analysis aimed at testing (mechanistic hypothesis on) disease progression and pharmacological interventions in patient-derived hiPSC-CMs.

【 授权许可】

Unknown