Parthenogenetic stem cells for tissue-engineered heart repair
Michael Didié, … , Loren J. Field, Wolfram-Hubertus Zimmermann
Michael Didié, … , Loren J. Field, Wolfram-Hubertus Zimmermann
Published February 22, 2013
Citation Information: J Clin Invest. 2013;123(3):1285-1298. https://doi.org/10.1172/JCI66854.
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Technical Advance

Parthenogenetic stem cells for tissue-engineered heart repair

Abstract

Uniparental parthenotes are considered an unwanted byproduct of in vitro fertilization. In utero parthenote development is severely compromised by defective organogenesis and in particular by defective cardiogenesis. Although developmentally compromised, apparently pluripotent stem cells can be derived from parthenogenetic blastocysts. Here we hypothesized that nonembryonic parthenogenetic stem cells (PSCs) can be directed toward the cardiac lineage and applied to tissue-engineered heart repair. We first confirmed similar fundamental properties in murine PSCs and embryonic stem cells (ESCs), despite notable differences in genetic (allelic variability) and epigenetic (differential imprinting) characteristics. Haploidentity of major histocompatibility complexes (MHCs) in PSCs is particularly attractive for allogeneic cell-based therapies. Accordingly, we confirmed acceptance of PSCs in MHC-matched allotransplantation. Cardiomyocyte derivation from PSCs and ESCs was equally effective. The use of cardiomyocyte-restricted GFP enabled cell sorting and documentation of advanced structural and functional maturation in vitro and in vivo. This included seamless electrical integration of PSC-derived cardiomyocytes into recipient myocardium. Finally, we enriched cardiomyocytes to facilitate engineering of force-generating myocardium and demonstrated the utility of this technique in enhancing regional myocardial function after myocardial infarction. Collectively, our data demonstrate pluripotency, with unrestricted cardiogenicity in PSCs, and introduce this unique cell type as an attractive source for tissue-engineered heart repair.

Authors

Michael Didié, Peter Christalla, Michael Rubart, Vijayakumar Muppala, Stephan Döker, Bernhard Unsöld, Ali El-Armouche, Thomas Rau, Thomas Eschenhagen, Alexander P. Schwoerer, Heimo Ehmke, Udo Schumacher, Sigrid Fuchs, Claudia Lange, Alexander Becker, Wen Tao, John A. Scherschel, Mark H. Soonpaa, Tao Yang, Qiong Lin, Martin Zenke, Dong-Wook Han, Hans R. Schöler, Cornelia Rudolph, Doris Steinemann, Brigitte Schlegelberger, Steve Kattman, Alec Witty, Gordon Keller, Loren J. Field, Wolfram-Hubertus Zimmermann

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Figure 6

Cardiomyocyte differentiation from PSCs in vitro and in vivo.

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Cardiomyocyte differentiation from PSCs in vitro and in vivo. (A) FACS o...
(A) FACS of GFP-positive PCMs (culture day 7 + 15); nontransgenic ESCs served as the control cell population (right panel illustrates a PCM 24 hours after plating; also refer to Supplemental Video 1). (B) Morphology of FACS-purified PCMs (EGP-positive) assessed by confocal laser scanning microscopy. (C) Distinct APs in FACS-purified PCMs. (D) Epifluorescence images and histochemical staining of atrial, atrioventricular (AV) nodal (acetylcholinesterase-positive/connexin43-negative), and ventricular regions in chimeric adult mouse heart (generated by blastocyst injection of αMHC-EGFP PSCs, A3 line). (E) Heart section from a chimeric mouse after injection of αMHC-EGFP PSCs (A3 line) into αMHC-nLacZ blastocysts. Light microscopy after X-gal staining (left panel); epifluorescence image of the same section showing PSC-derived EGFP-positive cardiomyocytes (middle panel); merged image did not show cells double-positive for EGFP and nLacZ, ruling out overt fusion events (right panel). Scale bars: (A) 10 μm, (B and D) 20 μm, (E) 50 μm.