Universität zu Köln

Lepperhof, Vera, Polchynski, Olga, Kruttwig, Klaus, Brueggemann, Chantal, Neef, Klaus, Drey, Florian, Zheng, Yunjie, Ackermann, Justus P., Choi, Yeong-Hoon, Wunderlich, Thomas F., Hoehn, Mathias, Hescheler, Juergen and Saric, Tomo ORCID: 0000-0001-8344-1095 (2014). Bioluminescent Imaging of Genetically Selected Induced Pluripotent Stem Cell-Derived Cardiomyocytes after Transplantation into Infarcted Heart of Syngeneic Recipients. PLoS One, 9 (9). SAN FRANCISCO: PUBLIC LIBRARY SCIENCE. ISSN 1932-6203

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Abstract

Cell loss after transplantation is a major limitation for cell replacement approaches in regenerative medicine. To assess the survival kinetics of induced pluripotent stem cell (iPSC)-derived cardiomyocytes (CM) we generated transgenic murine iPSC lines which, in addition to CM-specific expression of puromycin N-acetyl-transferase and enhanced green fluorescent protein (EGFP), also constitutively express firefly luciferase (FLuc) for bioluminescence (BL) in vivo imaging. While undifferentiated iPSC lines generated by random integration of the transgene into the genome retained stable FLuc activity over many passages, the BL signal intensity was strongly decreased in purified iPS-CM compared to undifferentiated iPSC. Targeted integration of FLuc-expression cassette into the ROSA26 genomic locus using zinc finger nuclease (ZFN) technology strongly reduced transgene silencing in iPS-CM, leading to a several-fold higher BL compared to iPS-CM expressing FLuc from random genomic loci. To investigate the survival kinetics of iPS-CM in vivo, purified CM obtained from iPSC lines expressing FLuc from a random or the ROSA26 locus were transplanted into cryoinfarcted hearts of syngeneic mice. Engraftment of viable cells was monitored by BL imaging over 4 weeks. Transplanted iPS-CM were poorly retained in the myocardium independently of the cell line used. However, up to 8% of cells survived for 28 days at the site of injection, which was confirmed by immunohistological detection of EGFP-positive iPS-CM in the host tissue. Transplantation of iPS-CM did not affect the scar formation or capillary density in the periinfarct region of host myocardium. This report is the first to determine the survival kinetics of drug-selected iPS-CM in the infarcted heart using BL imaging and demonstrates that transgene silencing in the course of iPSC differentiation can be greatly reduced by employing genome editing technology. FLuc-expressing iPS-CM generated in this study will enable further studies to reduce their loss, increase long-term survival and functional integration upon transplantation.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Lepperhof, Vera UNSPECIFIED UNSPECIFIED UNSPECIFIED Polchynski, Olga UNSPECIFIED UNSPECIFIED UNSPECIFIED Kruttwig, Klaus UNSPECIFIED UNSPECIFIED UNSPECIFIED Brueggemann, Chantal UNSPECIFIED UNSPECIFIED UNSPECIFIED Neef, Klaus UNSPECIFIED UNSPECIFIED UNSPECIFIED Drey, Florian UNSPECIFIED UNSPECIFIED UNSPECIFIED Zheng, Yunjie UNSPECIFIED UNSPECIFIED UNSPECIFIED Ackermann, Justus P. UNSPECIFIED UNSPECIFIED UNSPECIFIED Choi, Yeong-Hoon UNSPECIFIED UNSPECIFIED UNSPECIFIED Wunderlich, Thomas F. UNSPECIFIED UNSPECIFIED UNSPECIFIED Hoehn, Mathias UNSPECIFIED UNSPECIFIED UNSPECIFIED Hescheler, Juergen UNSPECIFIED UNSPECIFIED UNSPECIFIED Saric, Tomo UNSPECIFIED orcid.org/0000-0001-8344-1095 UNSPECIFIED
URN:	urn:nbn:de:hbz:38-428600
DOI:	10.1371/journal.pone.0107363
Journal or Publication Title:	PLoS One
Volume:	9
Number:	9
Date:	2014
Publisher:	PUBLIC LIBRARY SCIENCE
Place of Publication:	SAN FRANCISCO
ISSN:	1932-6203
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language ACUTE MYOCARDIAL-INFARCTION; IN-VIVO BIOLUMINESCENCE; COLLAGEN MATRICES; PROGENITOR CELLS; CARDIAC-FUNCTION; REPORTER GENES; SURVIVAL; ENGRAFTMENT; EXPRESSION; FIBROBLASTS Multiple languages Multidisciplinary Sciences Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/42860