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Extracellular Matrix-Mediated Maturation of Human Pluripotent Stem Cell-Derived Cardiac Monolayer Structure and Electrophysiological Function. | LitMetric

Extracellular Matrix-Mediated Maturation of Human Pluripotent Stem Cell-Derived Cardiac Monolayer Structure and Electrophysiological Function.

Circ Arrhythm Electrophysiol

From the Center for Arrhythmia Research, Department of Internal Medicine, Cardiovascular Research Center, University of Michigan, Ann Arbor, MI (T.J.H., A.M.D.R., K.C., D.P.-B., B.C.W., G.G.-S., Q.L., J.F., J.A., S.M., J.J.); Department of Medicine, University of California San Diego, La Jolla, CA (M.K.); Davis Heart and Lung Research Institute, Ohio State University, Columbus, OH (H.M.); Shanxi Medical University, Zhejiang, China (Q.L.); and University of Valencia, Valencia, Spain (M.Z., A.B.).

Published: April 2016

AI Article Synopsis

Article Abstract

Background: Human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) monolayers generated to date display an immature embryonic-like functional and structural phenotype that limits their utility for research and cardiac regeneration. In particular, the electrophysiological function of hPSC-CM monolayers and bioengineered constructs used to date are characterized by slow electric impulse propagation velocity and immature action potential profiles.

Methods And Results: Here, we have identified an optimal extracellular matrix for significant electrophysiological and structural maturation of hPSC-CM monolayers. hPSC-CM plated in the optimal extracellular matrix combination have impulse propagation velocities ≈2× faster than previously reported (43.6±7.0 cm/s; n=9) and have mature cardiomyocyte action potential profiles, including hyperpolarized diastolic potential and rapid action potential upstroke velocity (146.5±17.7 V/s; n=5 monolayers). In addition, the optimal extracellular matrix promoted hypertrophic growth of cardiomyocytes and the expression of key mature sarcolemmal (SCN5A, Kir2.1, and connexin43) and myofilament markers (cardiac troponin I). The maturation process reported here relies on activation of integrin signaling pathways: neutralization of β1 integrin receptors via blocking antibodies and pharmacological blockade of focal adhesion kinase activation prevented structural maturation.

Conclusions: Maturation of human stem cell-derived cardiomyocyte monolayers is achieved in a 1-week period by plating cardiomyocytes on PDMS (polydimethylsiloxane) coverslips rather than on conventional 2-dimensional cell culture formats, such as glass coverslips or plastic dishes. Activation of integrin signaling and focal adhesion kinase is essential for significant maturation of human cardiac monolayers.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4833010PMC
http://dx.doi.org/10.1161/CIRCEP.113.003638DOI Listing

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