Heparin Promotes Cardiac Differentiation of Human Pluripotent Stem Cells in Chemically Defined Albumin-Free Medium, Enabling Consistent Manufacture of Cardiomyocytes

Abstract

Cardiomyocytes can be differentiated from human pluripotent stem cells (hPSCs) in defined conditions, but efficient and consistent cardiomyocyte differentiation often requires expensive reagents such as B27 supplement or recombinant albumin. Using a chemically defined albumin‐free (E8 basal) medium, we identified heparin as a novel factor that significantly promotes cardiomyocyte differentiation efficiency, and developed an efficient method to differentiate hPSCs into cardiomyocytes. The treatment with heparin helped cardiomyocyte differentiation consistently reach at least 80% purity (up to 95%) from more than 10 different hPSC lines in chemically defined Dulbecco's modified Eagle's medium/F‐12‐based medium on either Matrigel or defined matrices like vitronectin and Synthemax. One of heparin's main functions was to act as a Wnt modulator that helped promote robust and consistent cardiomyocyte production. Our study provides an efficient, reliable, and cost‐effective method for cardiomyocyte derivation from hPSCs that can be used for potential large‐scale drug screening, disease modeling, and future cellular therapies. Stem Cells Translational Medicine 2017;6:527–538

Functional comparison of beating cardiomyocytes differentiated from umbilical cord‐derived mesenchymal/stromal stem cells and human foreskin‐derived induced pluripotent stem cells

“…These data are consistent with those reported by Lin et al (2017) and Lian et al (2012). However, this process could not be carried out on Synthemax™-coated PCL nanofibers due to detachment of iPSCs upon prolonged culture.…”

Single-Cell RNA-Sequencing and Optical Electrophysiology of Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes Reveal Discordance Between Cardiac Subtype-Associated Gene Expression Patterns and Electrophysiological Phenotypes

“…Together, this led us to profile individual cells via ArcLight and scRNA-seq at early (D12) and later (D40) stages of differentiation, and thus evaluate the cells both before and after distinct cardiomyocyte subtypes might presumably be detected. Electrophysiological classification of cardiomyocyte subtypes has frequently been performed during this time frame, adding additional relevance to our study design [11,[24][25][26][27][28][29].…”

“…Having established the utility of the ArcLight system and determined a classification system for APs, electrophysiological changes were mapped over the course of long-term culture of hiPSC-CMs (*6 weeks of differentiation), a time frame during which a mixed population of cardiomyocyte subtypes has often been described [11,[24][25][26][27][28][29]. Cardiomyocytes at D16 or earlier had a lower AP amplitude ( Fig.…”

Endogenous IGF Signaling Directs Heterogeneous Mesoderm Differentiation in Human Embryonic Stem Cells

“…Although cardiomyocytes can be efficiently derived with the WNT inhibitor and other modulators (Batalov and Feinberg, 2015;Lin et al, 2017), there are still many areas for improvement in cardiac differentiation and maturation. The IGF regulates not only the WNT pathway, but also metabolism and other important cellular activities (Chitnis et al, 2008;Freund et al, 1993).…”

“…We are interested in the endogenous signals involved in the cell fate diversity during mesoderm differentiation, and we try to direct lineage-specific differentiation by modulating endogenous signaling pathways. We previously established an albumin-free, chemically defined differentiation procedure to induce cardiomyocytes (Lin et al, 2017), and we modified the procedure to establish a stimuli-free neutral condition, which was used to identify factors involved in the cell fate diversity during mesoderm differentiation ( Figure 1A). In this report, we show that insulin/IGFs exert temporal regulation on mesoderm cell fates.…”