Effect of Human Donor Cell Source on Differentiation and Function of Cardiac Induced Pluripotent Stem Cells

• Published in: Journal of the American College of Cardiology Vol. 64; no. 5; pp. 436 - 448
• Main Authors: Sanchez-Freire, Veronica, PhD, Lee, Andrew S., PhD, Hu, Shijun, PhD, Abilez, Oscar J., MD, Liang, Ping, PhD, Lan, Feng, PhD, Huber, Bruno C., MD, Ong, Sang-Ging, PhD, Hong, Wan Xing, MS, Huang, Mei, PhD, Wu, Joseph C., MD, PhD
• Format: Journal Article
• Language: English
• Published: New York, NY Elsevier 05.08.2014; Elsevier Limited
• Subjects: Biological and medical sciences; Cardiology; Cardiology. Vascular system; Cardiovascular; Cardiovascular disease; Cell Differentiation - genetics; Cell differentiation, maturation, development, hematopoiesis; Cell physiology; Cells, Cultured; Efficiency; Embryonic Stem Cells - cytology; Epigenetics; Fibroblasts - cytology; Fibroblasts - physiology; Fundamental and applied biological sciences. Psychology; Gene expression; Gene Expression Regulation, Developmental; Heart; Humans; Induced Pluripotent Stem Cells - cytology; Induced Pluripotent Stem Cells - physiology; Influence; Internal Medicine; Medical sciences; Methods; Microscopy; Molecular and cellular biology; Myocytes, Cardiac - cytology; Normal distribution; Rodents; Statistical analysis; Stem Cell Transplantation; Stem cells; Studies; Tissue Donors; United States > US; California; cardiac troponin T; pluripotent stem cells; myocardial infarction; fluorescence-activated cell sorting; FACS; epigenetic memory; cTnT; MI; EB; stem cell antigen; EC; Fib; smooth muscle cell; DNA methylation; endothelial cell; iPSC; MEA; SMC; cardiac differentiation; CM; cardiac progenitor cell; embryoid body; Sca; fibroblast; induced pluripotent stem cell; multielectrode array; cardiomyocyte; CPC; Heart; Suboptimal donor; Human; Cell function; Stem cell; Cardiovascular disease; Circulatory system; Cardiology; Cell differentiation; Source
• ISSN: 0735-1097; 1558-3597
• DOI: 10.1016/j.jacc.2014.04.056

Abstract Background Human-induced pluripotent stem cells (iPSCs) are a potentially unlimited source for generation of cardiomyocytes (iPSC-CMs). However, current protocols for iPSC-CM derivation face several challenges, including variability in somatic cell sources and inconsistencies in cardiac differentiation efficiency. Objectives This study aimed to assess the effect of epigenetic memory on differentiation and function of iPSC-CMs generated from somatic cell sources of cardiac versus noncardiac origins. Methods Cardiac progenitor cells (CPCs) and skin fibroblasts from the same donors were reprogrammed into iPSCs and differentiated into iPSC-CMs via embryoid body and monolayer-based differentiation protocols. Results Differentiation efficiency was found to be higher in CPC-derived iPSC-CMs (CPC-iPSC-CMs) than in fibroblast-derived iPSC-CMs (Fib-iPSC-CMs). Gene expression analysis during cardiac differentiation demonstrated up-regulation of cardiac transcription factors in CPC-iPSC-CMs, including NKX2-5 , MESP1 , ISL1 , HAND2 , MYOCD, MEF2C, and GATA4 . Epigenetic assessment revealed higher methylation in the promoter region of NKX2-5 in Fib-iPSC-CMs compared with CPC-iPSC-CMs. Epigenetic differences were found to dissipate with increased cell passaging, and a battery of in vitro assays revealed no significant differences in their morphological and electrophysiological properties at early passage. Finally, cell delivery into a small animal myocardial infarction model indicated that CPC-iPSC-CMs and Fib-iPSC-CMs possess comparable therapeutic capabilities in improving functional recovery in vivo. Conclusions This is the first study to compare differentiation of iPSC-CMs from human CPCs versus human fibroblasts from the same donors. The authors demonstrate that although epigenetic memory improves differentiation efficiency of cardiac versus noncardiac somatic cell sources in vitro, it does not contribute to improved functional outcome in vivo.