Reporter-Based Isolation of Induced Pluripotent Stem Cell- and Embryonic Stem Cell-Derived Cardiac Progenitors Reveals Limited Gene Expression Variance

Gladstone Institute of Cardiovascular Disease, University of California, San Francisco, 94158, USA.
Circulation Research (Impact Factor: 11.02). 08/2010; 107(3):340-7. DOI: 10.1161/CIRCRESAHA.109.215434
Source: PubMed


Induced pluripotent stem (iPS) cells can differentiate into multiple cell types, including cardiomyocytes and have tremendous potential for drug discovery and regenerative therapies. However, it is unknown how much variability exists between differentiated lineages from independent iPS cell lines and, specifically, how similar iPS cell-derived cardiomyocytes (iPS-CMs) are to embryonic stem (ES) cell-derived cardiomyocytes (ES-CMs).
We investigated how much variability exists between differentiated lineages from independent iPS cell lines and how similar iPS-CMs are to ES-CMs.
We generated mouse iPS cells in which expression of NKX2-5, an early cardiac transcription factor, was marked by transgenic green fluorescent protein (GFP). Isolation of iPS- and ES-derived NKX2-5-GFP(+) cardiac progenitor pools, marked by identical reporters, revealed unexpectedly high similarity in genome-wide mRNA expression levels. Furthermore, the variability between cardiac progenitors derived from independent iPS lines was minimal. The NKX2-5-GFP(+) iPS cells formed cardiomyocytes by numerous induction protocols and could survive upon transplantation into the infarcted mouse heart without formation of teratomas.
Despite the line-to-line variability of gene expression in the undifferentiated state of ES and iPS cells, the variance narrows significantly in lineage-specific iPS-derived cardiac progenitors, and these progenitor cells can be isolated and used for transplantation without generation of unwanted cell types.

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Available from: Bruce Conklin, Mar 26, 2014
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    • "Fibroblasts were transduced with retroviral vectors (Oct 3/4, Sox2, Klf4, and c-Myc) which converted them to a cell type that is highly similar to the ESC, called induced pluripotent stem cell (iPSC).[69] Thus iPSCs have typical ESC markers, proliferate indefinitely, and are able to differentiate in somatic cells of all three embryonic cell lineages, including cardiomyocytes.[70],[71] An advantage of iPSCs is that ethical issues of ES cells are irrelevant and that autologous cells can be created, preventing immunological rejection. "
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    • "However, direct comparisons of CSCs with other cell sources are needed in order to identify the most effective cell type. Recently, novel approaches based on adult cell reprogramming (induced pluripotent stem cells; iPS) have been developed [33]. Although extremely promising, these strategies are still associated with potential safety concerns. "
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    • "A recent report indicated no significant difference in the fundamental characteristics of cardiomyocytes differentiated from either ES or iPS cells [35–38]. However, the efficiency of cardiogenesis from ES and iPS cells is still too low and not sufficiently stable to realize the goals of cardiac regenerative medicine. "
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    ABSTRACT: Induced pluripotent stem (iPS) cells are generated by reprogramming human somatic cells through the forced expression of several embryonic stem (ES) cell-specific transcription factors. The potential of iPS cells is having a significant impact on regenerative medicine, with the promise of infinite self-renewal, differentiation into multiple cell types, and no problems concerning ethics or immunological rejection. Human iPS cells are currently generated by transgene introduction principally through viral vectors, which integrate into host genomes, although the associated risk of tumorigenesis is driving research into nonintegration methods. Techniques for pluripotent stem cell differentiation and purification to yield cardiomyocytes are also advancing constantly. Although there remain some unsolved problems, cardiomyocyte transplantation may be a reality in the future. After those problems will be solved, applications of human iPS cells in human cardiovascular regenerative medicine will be envisaged for the future. Furthermore, iPS cell technology has generated new human disease models using disease-specific cells. This paper summarizes the progress of iPS cell technology in cardiovascular research.
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