title: Effect of human donor cell source on differentiation and function of cardiac induced pluripotent stem cells.
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.
  
  
  OBJECTIVE
  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.
authors.affiliation: Department of Medicine, Division of Cardiology, Stanford University School of Medicine, Stanford, California; Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, California; Department of Radiology, Stanford University School of Medicine, Stanford, California; Institute of Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, California.
authors.forename: Veronica
authors.initials: V
authors.lastname: Sanchez-Freire
authors.affiliation: Department of Medicine, Division of Cardiology, Stanford University School of Medicine, Stanford, California; Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, California; Institute of Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, California.
authors.forename: Andrew S
authors.initials: AS
authors.lastname: Lee
authors.affiliation: Department of Medicine, Division of Cardiology, Stanford University School of Medicine, Stanford, California; Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, California; Institute of Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, California.
authors.forename: Shijun
authors.initials: S
authors.lastname: Hu
authors.affiliation: Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, California.
authors.forename: Oscar J
authors.initials: OJ
authors.lastname: Abilez
authors.affiliation: Department of Medicine, Division of Cardiology, Stanford University School of Medicine, Stanford, California; Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, California; Department of Radiology, Stanford University School of Medicine, Stanford, California.
authors.forename: Ping
authors.initials: P
authors.lastname: Liang
authors.affiliation: Department of Medicine, Division of Cardiology, Stanford University School of Medicine, Stanford, California; Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, California; Department of Radiology, Stanford University School of Medicine, Stanford, California.
authors.forename: Feng
authors.initials: F
authors.lastname: Lan
authors.affiliation: Department of Medicine, Division of Cardiology, Stanford University School of Medicine, Stanford, California; Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, California.
authors.forename: Bruno C
authors.initials: BC
authors.lastname: Huber
authors.affiliation: Department of Medicine, Division of Cardiology, Stanford University School of Medicine, Stanford, California; Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, California; Department of Radiology, Stanford University School of Medicine, Stanford, California.
authors.forename: Sang-Ging
authors.initials: SG
authors.lastname: Ong
authors.affiliation: Department of Medicine, Division of Cardiology, Stanford University School of Medicine, Stanford, California; Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, California; Institute of Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, California.
authors.forename: Wan Xing
authors.initials: WX
authors.lastname: Hong
authors.affiliation: Department of Medicine, Division of Cardiology, Stanford University School of Medicine, Stanford, California; Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, California; Department of Radiology, Stanford University School of Medicine, Stanford, California.
authors.forename: Mei
authors.initials: M
authors.lastname: Huang
authors.affiliation: Department of Medicine, Division of Cardiology, Stanford University School of Medicine, Stanford, California; Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, California; Department of Radiology, Stanford University School of Medicine, Stanford, California; Institute of Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, California. Electronic address: joewu@stanford.edu.
authors.forename: Joseph C
authors.initials: JC
authors.lastname: Wu
mesh_terms: D002454:Cell Differentiation / Q000235:genetics; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology; D018507:Gene Expression Regulation, Developmental; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000502:physiology*; D032383:Myocytes, Cardiac / Q000166:cytology*; D033581:Stem Cell Transplantation; D014019:Tissue Donors
keywords: DNA methylation; cardiac differentiation; epigenetic memory; pluripotent stem cells
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