Limits...
Cardiopoietic programming of embryonic stem cells for tumor-free heart repair.

Behfar A, Perez-Terzic C, Faustino RS, Arrell DK, Hodgson DM, Yamada S, Puceat M, Niederländer N, Alekseev AE, Zingman LV, Terzic A - J. Exp. Med. (2007)

Bottom Line: Here, the tumorigenic threat associated with embryonic stem cell transplantation was suppressed by cardiac-restricted transgenic expression of the reprogramming cytokine TNF-alpha, enhancing the cardiogenic competence of recipient heart.Characterized by a down-regulation of oncogenic markers, up-regulation, and nuclear translocation of cardiac transcription factors, this predetermined population yielded functional cardiomyocyte progeny.Thus, cardiopoietic programming establishes a strategy to hone stem cell pluripotency, offering a tumor-resistant approach for regeneration.

View Article: PubMed Central - PubMed

Affiliation: Marriott Heart Disease Research Program, Division of Cardiovascular Diseases, Department of Medicine, Mayo Clinic, Rochester, MN 55905, USA.

ABSTRACT
Embryonic stem cells have the distinct potential for tissue regeneration, including cardiac repair. Their propensity for multilineage differentiation carries, however, the liability of neoplastic growth, impeding therapeutic application. Here, the tumorigenic threat associated with embryonic stem cell transplantation was suppressed by cardiac-restricted transgenic expression of the reprogramming cytokine TNF-alpha, enhancing the cardiogenic competence of recipient heart. The in vivo aptitude of TNF-alpha to promote cardiac differentiation was recapitulated in embryoid bodies in vitro. The procardiogenic action required an intact endoderm and was mediated by secreted cardio-inductive signals. Resolved TNF-alpha-induced endoderm-derived factors, combined in a cocktail, secured guided differentiation of embryonic stem cells in monolayers produce cardiac progenitors termed cardiopoietic cells. Characterized by a down-regulation of oncogenic markers, up-regulation, and nuclear translocation of cardiac transcription factors, this predetermined population yielded functional cardiomyocyte progeny. Recruited cardiopoietic cells delivered in infarcted hearts generated cardiomyocytes that proliferated into scar tissue, integrating with host myocardium for tumor-free repair. Thus, cardiopoietic programming establishes a strategy to hone stem cell pluripotency, offering a tumor-resistant approach for regeneration.

Show MeSH

Related in: MedlinePlus

Cardiopoietic stem cells are confined to the cardiac program. (A) Hanging drop differentiation of embryonic stem cells resulted in differentiation of cells into all three germinal layers. (B) Treatment of embryonic stem cells with a single component (e.g., TGF-β1) of the cardiogenic cocktail increases stem cell aptitude for cardiogenesis—demonstrated by increased beating area—but still results in trigerminal embryoid body formation indicating maintained pluripotency. (C) Differentiation of cardiopoietic cells generated cardiospheres demonstrating definitive propensity toward cardiogenesis. (D–G) Isolation of cells from cardiopoietic cell–generated cardiospheres yielded functional cardiomyocytes demonstrating action potential (E), ion current activity (F), and calcium transients (G) along with organized sarcomerogenesis recapitulating myocyte structure on confocal and transmission electron microscopy (G, inset). Bars: (A–C) 500 μm; (F) 3 s; (G) 20 μm; (G, inset) 1 μm.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2118723&req=5

fig8: Cardiopoietic stem cells are confined to the cardiac program. (A) Hanging drop differentiation of embryonic stem cells resulted in differentiation of cells into all three germinal layers. (B) Treatment of embryonic stem cells with a single component (e.g., TGF-β1) of the cardiogenic cocktail increases stem cell aptitude for cardiogenesis—demonstrated by increased beating area—but still results in trigerminal embryoid body formation indicating maintained pluripotency. (C) Differentiation of cardiopoietic cells generated cardiospheres demonstrating definitive propensity toward cardiogenesis. (D–G) Isolation of cells from cardiopoietic cell–generated cardiospheres yielded functional cardiomyocytes demonstrating action potential (E), ion current activity (F), and calcium transients (G) along with organized sarcomerogenesis recapitulating myocyte structure on confocal and transmission electron microscopy (G, inset). Bars: (A–C) 500 μm; (F) 3 s; (G) 20 μm; (G, inset) 1 μm.

Mentions: Genomic expression profiling demonstrated progressive loss of pluripotent traits associated with cardiac specification of embryonic stem cells (Fig. 7 F). Dissection of the cardiopoietic transcriptome revealed down-regulation in markers of pluripotency (Oct4; reference 27) and oncogenicity (MYC and DEK; reference 49) along with activation of cardiogenic pathways (MEF-2C, Nkx2.5, GATA4, and Tbx2), preceding expression of the excitation-contraction machinery (L-type Ca2+ channel, MLC2v) typical of the cardiac lineage (Fig. 7 F). Differentiation of embryonic stem cells untreated or treated with single factors from the cardiogenic cocktail yielded three germinal layer embryoid bodies (Fig. 8, A and B), reflecting a maintained pluripotency. In contrast, day 4–recruited cardiopoietic cells no longer produced trigerminal layer embryoid bodies and instead formed beating cardiospheres, demonstrating commitment to the cardiac program (Fig. 8 C). Dissociation of cardiospheres released cells with hallmark features of cardiomyocytes, including action potential activity (Fig. 8 D), ion current profiles (Fig. 8 E), rhythmic calcium transients (Fig. 8 F), and sarcomeric organization (Fig. 8 G). Thus, derived cardiopoietic cells exchange the pluripotent and oncogenic molecular profile of embryonic stem cells for definitive commitment to cardiac lineage.


Cardiopoietic programming of embryonic stem cells for tumor-free heart repair.

Behfar A, Perez-Terzic C, Faustino RS, Arrell DK, Hodgson DM, Yamada S, Puceat M, Niederländer N, Alekseev AE, Zingman LV, Terzic A - J. Exp. Med. (2007)

Cardiopoietic stem cells are confined to the cardiac program. (A) Hanging drop differentiation of embryonic stem cells resulted in differentiation of cells into all three germinal layers. (B) Treatment of embryonic stem cells with a single component (e.g., TGF-β1) of the cardiogenic cocktail increases stem cell aptitude for cardiogenesis—demonstrated by increased beating area—but still results in trigerminal embryoid body formation indicating maintained pluripotency. (C) Differentiation of cardiopoietic cells generated cardiospheres demonstrating definitive propensity toward cardiogenesis. (D–G) Isolation of cells from cardiopoietic cell–generated cardiospheres yielded functional cardiomyocytes demonstrating action potential (E), ion current activity (F), and calcium transients (G) along with organized sarcomerogenesis recapitulating myocyte structure on confocal and transmission electron microscopy (G, inset). Bars: (A–C) 500 μm; (F) 3 s; (G) 20 μm; (G, inset) 1 μm.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2118723&req=5

fig8: Cardiopoietic stem cells are confined to the cardiac program. (A) Hanging drop differentiation of embryonic stem cells resulted in differentiation of cells into all three germinal layers. (B) Treatment of embryonic stem cells with a single component (e.g., TGF-β1) of the cardiogenic cocktail increases stem cell aptitude for cardiogenesis—demonstrated by increased beating area—but still results in trigerminal embryoid body formation indicating maintained pluripotency. (C) Differentiation of cardiopoietic cells generated cardiospheres demonstrating definitive propensity toward cardiogenesis. (D–G) Isolation of cells from cardiopoietic cell–generated cardiospheres yielded functional cardiomyocytes demonstrating action potential (E), ion current activity (F), and calcium transients (G) along with organized sarcomerogenesis recapitulating myocyte structure on confocal and transmission electron microscopy (G, inset). Bars: (A–C) 500 μm; (F) 3 s; (G) 20 μm; (G, inset) 1 μm.
Mentions: Genomic expression profiling demonstrated progressive loss of pluripotent traits associated with cardiac specification of embryonic stem cells (Fig. 7 F). Dissection of the cardiopoietic transcriptome revealed down-regulation in markers of pluripotency (Oct4; reference 27) and oncogenicity (MYC and DEK; reference 49) along with activation of cardiogenic pathways (MEF-2C, Nkx2.5, GATA4, and Tbx2), preceding expression of the excitation-contraction machinery (L-type Ca2+ channel, MLC2v) typical of the cardiac lineage (Fig. 7 F). Differentiation of embryonic stem cells untreated or treated with single factors from the cardiogenic cocktail yielded three germinal layer embryoid bodies (Fig. 8, A and B), reflecting a maintained pluripotency. In contrast, day 4–recruited cardiopoietic cells no longer produced trigerminal layer embryoid bodies and instead formed beating cardiospheres, demonstrating commitment to the cardiac program (Fig. 8 C). Dissociation of cardiospheres released cells with hallmark features of cardiomyocytes, including action potential activity (Fig. 8 D), ion current profiles (Fig. 8 E), rhythmic calcium transients (Fig. 8 F), and sarcomeric organization (Fig. 8 G). Thus, derived cardiopoietic cells exchange the pluripotent and oncogenic molecular profile of embryonic stem cells for definitive commitment to cardiac lineage.

Bottom Line: Here, the tumorigenic threat associated with embryonic stem cell transplantation was suppressed by cardiac-restricted transgenic expression of the reprogramming cytokine TNF-alpha, enhancing the cardiogenic competence of recipient heart.Characterized by a down-regulation of oncogenic markers, up-regulation, and nuclear translocation of cardiac transcription factors, this predetermined population yielded functional cardiomyocyte progeny.Thus, cardiopoietic programming establishes a strategy to hone stem cell pluripotency, offering a tumor-resistant approach for regeneration.

View Article: PubMed Central - PubMed

Affiliation: Marriott Heart Disease Research Program, Division of Cardiovascular Diseases, Department of Medicine, Mayo Clinic, Rochester, MN 55905, USA.

ABSTRACT
Embryonic stem cells have the distinct potential for tissue regeneration, including cardiac repair. Their propensity for multilineage differentiation carries, however, the liability of neoplastic growth, impeding therapeutic application. Here, the tumorigenic threat associated with embryonic stem cell transplantation was suppressed by cardiac-restricted transgenic expression of the reprogramming cytokine TNF-alpha, enhancing the cardiogenic competence of recipient heart. The in vivo aptitude of TNF-alpha to promote cardiac differentiation was recapitulated in embryoid bodies in vitro. The procardiogenic action required an intact endoderm and was mediated by secreted cardio-inductive signals. Resolved TNF-alpha-induced endoderm-derived factors, combined in a cocktail, secured guided differentiation of embryonic stem cells in monolayers produce cardiac progenitors termed cardiopoietic cells. Characterized by a down-regulation of oncogenic markers, up-regulation, and nuclear translocation of cardiac transcription factors, this predetermined population yielded functional cardiomyocyte progeny. Recruited cardiopoietic cells delivered in infarcted hearts generated cardiomyocytes that proliferated into scar tissue, integrating with host myocardium for tumor-free repair. Thus, cardiopoietic programming establishes a strategy to hone stem cell pluripotency, offering a tumor-resistant approach for regeneration.

Show MeSH
Related in: MedlinePlus