Nanotopography-Induced Structural Anisotropy and Sarcomere Development in Human Cardiomyocytes Derived from Induced Pluripotent Stem Cells

Abstract

Understanding the phenotypic development of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) is a prerequisite to advancing regenerative cardiac therapy, disease modeling, and drug screening applications. Lack of consistent hiPSC-CM in vitro data can be largely attributed to the inability of conventional culture methods to mimic the structural, biochemical, and mechanical aspects of the myocardial niche accurately. Here, we present a nanogrid culture array comprised of nanogrooved topographies, with groove widths ranging from 350 to 2000 nm, to study the effect of different nanoscale structures on the structural development of hiPSC-CMs in vitro. Nanotopographies were designed to have a biomimetic interface, based on observations of the oriented myocardial extracellular matrix (ECM) fibers found in vivo. Nanotopographic substrates were integrated with a self-assembling chimeric peptide containing the Arg-Gly-Asp (RGD) cell adhesion motif. Using this platform, cell adhesion to peptide-coated substrates was found to be comparable to that of conventional fibronectin-coated surfaces. Cardiomyocyte organization and structural development were found to be dependent on the nanotopographical feature size in a biphasic manner, with improved development achieved on grooves in the 700–1000 nm range. These findings highlight the capability of surface-functionalized, bioinspired substrates to influence cardiomyocyte development, and the capacity for such platforms to serve as a versatile assay for investigating the role of topographical guidance cues on cell behavior. Such substrates could potentially create more physiologically relevant in vitro cardiac tissues for future drug screening and disease modeling studies.

Human iPSC-derived cardiomyocytes and tissue engineering strategies for disease modeling and drug screening

“…One example of an attempt to mimic the native myocardial niche in vitro is the utilization of biomimetic nano-structured surfaces to improve cell adhesion, proliferation, and migration, as well as cardiomyogenic differentiation and structural development (Carson et al, 2016, Kim, Kshitiz, 2012a). Parallel nano-grooves and nano-ridges help maintain the cell polarity and alignment important for organ development by providing contact guidance which influences the organization of microtubules, focal contacts, and actin filaments in parallel with the underlying topography (Kim et al, 2012b).…”

Multiscale technologies for treatment of ischemic cardiomyopathy

“…In addition, nanopatterned substrates have been widely used to encourage maturation of chemically defined CMs 73–76 . However, new methods proposed thus far have not yielded highly mature, patient-specific CMs on a scale sufficient to meet clinical needs 77 .…”

Advances and Current Challenges Associated with the Use of Human Induced Pluripotent Stem Cells in Modeling Neurodegenerative Disease

“…Such substrates enable control of neuritic outgrowth and polarity, which may also offer benefits when seeking to develop controlled neuronal networks [Ferrari et al, 2011]. Studies into other electrically active cell types, like cardiomyocytes, have demonstrated that topographic patterning can enhance physiological structure and function [Yang et al, 2014a;Carson et al, 2016] but whether or not such surfaces enhance network connectivity and functional performance in iPSC-based neuronal cultures remains to be seen. Highly conductive substrates have been shown to improve network activity in cultured neuronal populations [Lovat et al, 2005;Tang et al, 2013].…”