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PMID: 21494607 Published · epublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

A universal system for highly efficient cardiac differentiation of human induced pluripotent stem cells that eliminates interline variability.

PloS one ·Vol. 6 ·No. 4 ·2011-04-08 ·Pages e18293

Burridge PW, Thompson S, Millrod MA, Weinberg S, Yuan X, Peters A, Mahairaki V, Koliatsos VE, Tung L, Zambidis ET

Abstract

The production of cardiomyocytes from human induced pluripotent stem cells (hiPSC) holds great promise for patient-specific cardiotoxicity drug testing, disease modeling, and cardiac regeneration. However, existing protocols for the differentiation of hiPSC to the cardiac lineage are inefficient and highly variable. We describe a highly efficient system for differentiation of human embryonic stem cells (hESC) and hiPSC to the cardiac lineage. This system eliminated the variability in cardiac differentiation capacity of a variety of human pluripotent stem cells (hPSC), including hiPSC generated from CD34(+) cord blood using non-viral, non-integrating methods. We systematically and rigorously optimized >45 experimental variables to develop a universal cardiac differentiation system that produced contracting human embryoid bodies (hEB) with an improved efficiency of 94.7±2.4% in an accelerated nine days from four hESC and seven hiPSC lines tested, including hiPSC derived from neonatal CD34(+) cord blood and adult fibroblasts using non-integrating episomal plasmids. This cost-effective differentiation method employed forced aggregation hEB formation in a chemically defined medium, along with staged exposure to physiological (5%) oxygen, and optimized concentrations of mesodermal morphogens BMP4 and FGF2, polyvinyl alcohol, serum, and insulin. The contracting hEB derived using these methods were composed of high percentages (64-89%) of cardiac troponin I(+) cells that displayed ultrastructural properties of functional cardiomyocytes and uniform electrophysiological profiles responsive to cardioactive drugs. This efficient and cost-effective universal system for cardiac differentiation of hiPSC allows a potentially unlimited production of functional cardiomyocytes suitable for application to hPSC-based drug development, cardiac disease modeling, and the future generation of clinically-safe nonviral human cardiac cells for regenerative medicine.

MeSH Terms
Adult Animals Antigens, CD34/metabolism Body Patterning/drug effects Bone Morphogenetic Protein 4/pharmacology Cell Adhesion/drug effects Cell Culture Techniques/methods Cell Differentiation/drug effects Cell Line Cell Proliferation/drug effects Culture Media/pharmacology Electrophysiological Phenomena/drug effects Embryoid Bodies/cytology,drug effects,metabolism Fetal Blood/cytology Fibroblast Growth Factor 2/pharmacology Fibroblasts/cytology,drug effects,metabolism Genetic Vectors/genetics Humans Induced Pluripotent Stem Cells/cytology,drug effects,metabolism Insulin/pharmacology Mesoderm/cytology,drug effects Mice Myocytes, Cardiac/cytology,drug effects,metabolism Oxygen/pharmacology Polyvinyl Alcohol/pharmacology Transgenes/genetics
Chemicals
Antigens, CD34 Bone Morphogenetic Protein 4 Culture Media Insulin Fibroblast Growth Factor 2 Polyvinyl Alcohol Oxygen
Authors & Affiliations
10 authors, click to expand affiliations / ORCID
Burridge Paul W
Johns Hopkins Institute for Cell Engineering, The Johns Hopkins University School of Medicine, Baltimore, Maryland, United States of America. [email protected]
Thompson Susan
Millrod Michal A
Weinberg Seth
Yuan Xuan
Peters Ann
Mahairaki Vasiliki
Koliatsos Vassilis E
Tung Leslie
Zambidis Elias T
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Article Info
Journal
PloS one
Abbr.
PLoS One
ISSN
1932-6203
DOI
10.1371/journal.pone.0018293
Email
[email protected]
Published
2011-04-08
Epub
2011-00-08
Pages
e18293
Language
English
Region
United States
NLM ID
101285081
PMCID
PMC3072973
Subset
IM
Grants
NHLBI NIH HHS · U01 HL100397 · United States
NHLBI NIH HHS · U01 HL099775 · United States
NHLBI NIH HHS · R03 HL096220 · United States
NHLBI NIH HHS · U01HL099775 · United States
NHLBI NIH HHS · U01HL100397 · United States
NIAID NIH HHS · T32 AI007247 · United States
NHLBI NIH HHS · K08 HL077595 · United States
Corrections
CommentIn
PMID: 21985539
External Links
PubMed source DOI: 10.1371/journal.pone.0018293 PMC Full Text View journal details
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