title: Enhanced Human-Induced Pluripotent Stem Cell Derived Cardiomyocyte Maturation Using a Dual Microgradient Substrate.
abstract: Human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) raise many possibilities for cardiac research but they exhibit an immature phenotype, which influences experimental outcomes. The aim of our research is to investigate the effects of a topographical gradient substrate on the morphology and function of commercially available hiPSC-CM. The lateral dimensions the microgrooves on the substrate varied from 8 to 100 μm space between the 8 μm grooves on one axis and from ∼5 nm to ∼1 μm in depth on the other axis. Cells were seeded homogeneously across the substrate and according to the manufacturers protocols. At days 4 and 10, measures of eccentricity, elongation, orientation, sarcomere length (SL), and contractility of the hiPSC-CM were taken. Only the deepest and widest region (8-30 μm wide and 0.85-1 μm deep) showed a significantly higher percentage of hiPSC-CM with an increased eccentricity (31.3 ± 6.4%), elongation (10.4 ± 4.3%), and orientation (<10°) (32.1 ± 2.7%) when compared with the control (flat substrate) (15.8 ± 5.0%, 3.4 ± 2.7%, and 10.6 ± 1.1%, respectively). Additionally, during stimulus-induced contraction, the relaxation phase of the twitch was prolonged (400 ms) compared to nonelongated cells (200 ms). These findings support the potential use of dual microgradient substrates to investigate substrate topographies that stimulate migration and/or maturation of hiPSC-CM.
authors.affiliation: Institute of Cardiovascular and Medical Sciences, University of Glasgow, Glasgow G12 8QQ, United Kingdom; Division of Biomedical Engineering, School of Engineering, University of Glasgow, Glasgow G12 8LT, United Kingdom.
authors.forename: E
authors.initials: E
authors.lastname: Huethorst
authors.affiliation: Institute of Cardiovascular and Medical Sciences, University of Glasgow , Glasgow G12 8QQ, United Kingdom.
authors.forename: M
authors.initials: M
authors.lastname: Hortigon
authors.affiliation: Institute of Cardiovascular and Medical Sciences, University of Glasgow , Glasgow G12 8QQ, United Kingdom.
authors.forename: V
authors.initials: V
authors.lastname: Zamora-Rodriguez
authors.affiliation: Division of Biomedical Engineering, School of Engineering, University of Glasgow , Glasgow G12 8LT, United Kingdom.
authors.forename: P M
authors.initials: PM
authors.lastname: Reynolds
authors.affiliation: Institute of Cardiovascular and Medical Sciences, University of Glasgow , Glasgow G12 8QQ, United Kingdom.
authors.forename: F
authors.initials: F
authors.lastname: Burton
authors.affiliation: Institute of Cardiovascular and Medical Sciences, University of Glasgow , Glasgow G12 8QQ, United Kingdom.
authors.forename: G
authors.initials: G
authors.lastname: Smith
authors.affiliation: Division of Biomedical Engineering, School of Engineering, University of Glasgow , Glasgow G12 8LT, United Kingdom.
authors.forename: N
authors.initials: N
authors.lastname: Gadegaard
keywords: biomaterial; cardiac regeneration; functionality; hiPSC-CM; microgrooves; topography