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

Year: 2019

Authors: Pioner JM., Santini L., Palandri C., Martella D., Lupi F., Langione M., Querceto S., Grandinetti B., Balducci V., Benzoni P., Landi S., Barbuti A., Lupi FF., Boarino L., Sartiani L., Tesi C., Mack DL., Regnier M., Cerbai E., Parmeggiani C., Poggesi C., Ferrantini C., Coppini R.

Autors Affiliation: Univ Firenze, Dept Expt & Clin Med, Div Physiol, I-50134 Florence, Italy; Univ Florence, Dept NeuroFarBa, I-50134 Florence, Italy; European Lab Nonlinear Spect LENS, I-50019 Florence, Italy; CNR, INO, Natl Inst Opt, I-50125 Florence, Italy; Univ Milan, Dept Biosci, I-20137 Milan, Italy; Ist Nazl Ric Metrol INRiM, I-10129 Turin, Italy; Univ Washington, Dept Rehabil Med, Seattle, WA 98108 USA; Univ Washington, Dept Bioengn, Seattle, WA 98108 USA; Univ Florence, Dept Chem Ugo Schiff, I-50134 Florence, Italy.

Abstract: 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 beta-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.

Journal/Review: INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES

Volume: 20 (15)      Pages from: 3799-1  to: 3799-17

More Information: This research received fundings from: Telethon Italy under grant agreement No GGP16191 (C.F.); the European Union?s Horizon 2020 research and innovation program under grant agreement No 777204 (C.P.); Ente Cassa di Risparmio di Firenze (grant 2017/0713) (E.C.).
KeyWords: human induced pluripotent stem cells; cardiomyocytes; fluorescence; maturation; action potential; calcium handling; hydrogels; long-term culture
DOI: 10.3390/ijms20153799

Citations: 29
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