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Lineage tracing of cardiac explant derived cells.

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journal contribution
posted on 24.10.2012, 08:58 by Lincoln T. Shenje, L. J. Field, Catrin A. Pritchard, Christopher J. Guerin, M. Rubart, M. H. Soonpaa, Keng-Leong Ang, Manuel Galiñanes
Aims Cultured cardiac explants produce a heterogeneous population of cells including a distinctive population of refractile cells described here as small round cardiac explant derived cells (EDCs). The aim of this study was to explore the source, morphology and cardiogenic potential of EDCs. Methods Transgenic MLC2v-Cre/ZEG, and actin-eGFP mice were used for lineage-tracing of EDCs in vitro and in vivo. C57B16 mice were used as cell transplant recipients of EDCs from transgenic hearts, as well as for the general characterisation of EDCs. The activation of cardiac-specific markers were analysed by: immunohistochemistry with bright field and immunofluorescent microscopy, electron microscopy, PCR and RT-PCR. Functional engraftment of transplanted cells was further investigated with calcium transient studies. Results Production of EDCs was highly dependent on the retention of blood-derived cells or factors in the cultured explants. These cells shared some characteristics of cardiac myocytes in vitro and survived engraftment in the adult heart in vivo. However, EDCs failed to differentiate into functional cardiac myocytes in vivo as demonstrated by the absence of stimulation-evoked intracellular calcium transients following transplantation into the peri-infarct zone. Conclusions This study highlights that positive identification based upon one parameter alone such as morphology or immunofluorescene is not adequate to identify the source, fate and function of adult cardiac explant derived cells.

Funding

Bristol Myer Squib Cardiovascular Prize Fellowship 2003 (LTS)

History

Citation

PLoS ONE, 2008, 3 (4), pp. e1929-e1929

Published in

PLoS ONE

Publisher

Public Library of Science

issn

1932-6203

eissn

1932-6203

Copyright date

2008

Available date

24/10/2012

Publisher version

http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0001929

Language

eng