Cardiomyocytes From Human and Mouse Embryonic Stem Cells

Abstract

SummaryHuman and mouse embryonic stem (ES) cells have the potential to differentiate to cardiomyocytes in culture. They are therefore of interest for studying early human and mouse heart development, as well as properties of cardiomyocytes from both species, including their responses to cardiac drugs, and, at some point in the future, may represent a source of transplantable cells for cardiac muscle repair. The differentiation protocols that are effective depend in part on the species from which the ES cell lines were derived, and in part on the individual cell lines and the methods used for their propagation prior to differentiation. Here, several methods for generating and characterizing cardiomyocytes from mouse and human ES cells are described, as well as methods for dissociation of cardiomyocytes into single-cell suspensions which are useful both for characterizing cells by antibody staining and electrophysiological measurements, as well as preparing cells for transplantation into (animal) hearts.

Cardiomyocytes Derived From Pluripotent Stem Cells Recapitulate Electrophysiological Characteristics of an Overlap Syndrome of Cardiac Sodium Channel Disease

“…27,28 Teratomas were generated by injecting undifferentiated iPSCs into nonobese diabetic severe combined immunodeficiency (NOD-SCID) mice. mPSCs were differentiated in vitro as embryoid bodies (EBs) using a "hanging drop" protocol, 27 whereas hiPSCs were induced to differentiate to cardiomyocytes by coculture on visceral endoderm-like (END-2) cells as described previously. 28,29 Immunofluorescence and gene expression analyses were performed as described previously.…”

“…27,31 One day after EBs were transferred to adherent culture (day 8 of differentiation), spontaneously contracting foci were apparent (Movie I in the online-only Data Supplement). By day 10 of differentiation, 75% to 90% of mESC-and miPSC-derived EBs contained spontaneously contracting regions, indicating comparable cardiomyocyte differentiation potentials ( Figure IIA in the online-only Data Supplement).…”

SK4 Ca 2+ activated K + channel is a critical player in cardiac pacemaker derived from human embryonic stem cells

“…After 7 d of culture in suspension, EBs were plated on 0.1% gelatin-coated plates and checked daily until a spontaneous beating activity was visible (generally onset of beating at day 9 after the initiation of differentiation). Beating clusters were mechanically dissected from EBs, following a three-step dissociation protocol as previously described (54). The hESC-CMs were isolated and plated on Matrigel or 0.1% fibronectin-coated glass coverslips (13 mm diameter) in 24-well plates.…”

Developmental and Functional Nature of Human iPSC Derived Motoneurons

“…Directed differentiation is influenced by co-culture with different somatic cells [15,137] and/or by supplementation of growth factors and small molecules [106,213]. Addition of rock-inhibitor (rho-associated kinase inhibitor) Y-27632 for the first 24h improves survival of human ES single cells and promotes generation of EBs [63].…”