A purified population of multipotent cardiovascular progenitors derived from primate pluripotent stem cells engrafts in postmyocardial infarcted nonhuman primates
Guillaume Blin, … , Philippe Menasché, Michel Pucéat
Guillaume Blin, … , Philippe Menasché, Michel Pucéat
Published March 24, 2010
Citation Information: J Clin Invest. 2010;120(4):1125-1139. https://doi.org/10.1172/JCI40120.
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Research Article

A purified population of multipotent cardiovascular progenitors derived from primate pluripotent stem cells engrafts in postmyocardial infarcted nonhuman primates

Abstract

Cell therapy holds promise for tissue regeneration, including in individuals with advanced heart failure. However, treatment of heart disease with bone marrow cells and skeletal muscle progenitors has had only marginal positive benefits in clinical trials, perhaps because adult stem cells have limited plasticity. The identification, among human pluripotent stem cells, of early cardiovascular cell progenitors required for the development of the first cardiac lineage would shed light on human cardiogenesis and might pave the way for cell therapy for cardiac degenerative diseases. Here, we report the isolation of an early population of cardiovascular progenitors, characterized by expression of OCT4, stage-specific embryonic antigen 1 (SSEA-1), and mesoderm posterior 1 (MESP1), derived from human pluripotent stem cells treated with the cardiogenic morphogen BMP2. This progenitor population was multipotential and able to generate cardiomyocytes as well as smooth muscle and endothelial cells. When transplanted into the infarcted myocardium of immunosuppressed nonhuman primates, an SSEA-1+ progenitor population derived from Rhesus embryonic stem cells differentiated into ventricular myocytes and reconstituted 20% of the scar tissue. Notably, primates transplanted with an unpurified population of cardiac-committed cells, which included SSEA-1– cells, developed teratomas in the scar tissue, whereas those transplanted with purified SSEA-1+ cells did not. We therefore believe that the SSEA-1+ progenitors that we have described here have the potential to be used in cardiac regenerative medicine.

Authors

Guillaume Blin, David Nury, Sonia Stefanovic, Tui Neri, Oriane Guillevic, Benjamin Brinon, Valérie Bellamy, Catherine Rücker-Martin, Pascal Barbry, Alain Bel, Patrick Bruneval, Chad Cowan, Julia Pouly, Shoukhrat Mitalipov, Elodie Gouadon, Patrice Binder, Albert Hagège, Michel Desnos, Jean-François Renaud, Philippe Menasché, Michel Pucéat

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Figure 1

Gene and protein profiles of SSEA-1+ cardiac progenitors.

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Gene and protein profiles of SSEA-1+ cardiac progenitors. ESCs were ...
ESCs were treated or not (CTRL) for 4 days with BMP2 (10 ng/ml) and (A) monitored by flow cytometry using a FITC-conjugated anti–SSEA-1 antibody or (B and C) separated using the anti–SSEA-1 Miltenyi kit and the MACS columns. SSEA-1– and SSEA-1+ cDNAs were run in real-time PCR. Data are from 8 experiments, performed on the HUES-24 cell line, and reproduced in different HUESC lines (H9, HUES-9, HUES-24, HUES-26, I3, and I6). The inset in C illustrates SSEA-1+ cells immunostained with anti-Tbx6 and anti-Mesp1/2 antibody 12–24 hours after sorting. (D and E) ChIP assay using (D) anti-H3triMeK4 or anti-triMeK27 antibodies or (E) the anti-PolII or serine-phosphorylated PolII (Ser5-PPolII, left) antibodies. Q-PCR was used to amplify the chromatin-bound DNA, using primers specific for OCT4, TBX6, ISL1, MEF2C, NKX2.5, ACTC1, and PAX4 promoters (sequence within the 700 bp in 3′ of ATG; Table 2). Data (n = 5) show fold enrichment of methylated histones on promoters in the SSEA-1+ versus SSEA-1– population. The gels illustrate the specificity of PCR products (input, SSEA-1+ DNA samples from anti-triMeH3K4 IP, anti-rabbit IgG, or no antibody). OCT4IA, OCT-4 isoform A. (E) The left side shows PCR gel of DNA products after real-time PCR and indicates a complete loss of serine-phosphorylated PolII on both the NANOG and SOX2 promoters. ChiP experiments have been mostly performed using the HUES-24 cell line and were validated in 2 other experiments using the I6 cell line. *P ≤ 0.05; #P ≤ 0.01.