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 5

Mesodermal differentiation and cardiogenicity.

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Mesodermal differentiation and cardiogenicity. qPCR for pluripotency (Po...
qPCR for pluripotency (Pou5f1), (cardio)mesoderm (T, Kdr, Isl1), and cardiac (Nkx2-5, Myh6) markers in undifferentiated (UD) cells and differentiating EBs in (A) PSC (n = 4–5 per group) and (B) ESC (n = 3–4 per group) cultures (data represent means ± SEM). Decrease in Nkx2-5 and Mhy6 in ESCs on culture day 7 + 6 was the consequence of nonmyocyte overgrowth. (C) Enumeration of beating EBs at the indicated culture days (n = 2–3 per time point; data represent means). (D) Quantity of EGFP-positive cardiomyocytes (CMs) per input stem cell (SC; 400 ESCs or PSCs per EB; cells from 5 single EBs were pooled for analytic FACS; n = 5–6) on culture day 5 + 10 (box plots indicate median with interquartile range; whiskers indicate minimal and maximal values). PSC line A3 and ESC lines ES4 and ES7 were generated from the same αMHC-EGFP mouse strain.