Lineage tracing of cardiac explant derived cells

Lincoln T. Shenje, Loren J. Field, Catrin A. Pritchard, Christopher J. Guerin, Michael Rubart, Mark H. Soonpaa, Keng Leong Ang, Manuel Galiñanes

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

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 immunofluorescence is not adequate to identify the source, fate and function of adult cardiac explant derived cells.

Original languageEnglish (US)
Article numbere1929
JournalPLoS One
Volume3
Issue number4
DOIs
StatePublished - Apr 16 2008
Externally publishedYes

Fingerprint

explants
Calcium
Transplants
Electron microscopy
Actins
Blood
Chemical activation
Cells
cells
Cardiac Myocytes
heart
Polymerase Chain Reaction
genetically modified organisms
calcium
infarction
mice
Population
Fluorescent Antibody Technique
Immunohistochemistry
Microscopy

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Shenje, L. T., Field, L. J., Pritchard, C. A., Guerin, C. J., Rubart, M., Soonpaa, M. H., ... Galiñanes, M. (2008). Lineage tracing of cardiac explant derived cells. PLoS One, 3(4), [e1929]. https://doi.org/10.1371/journal.pone.0001929

Lineage tracing of cardiac explant derived cells. / Shenje, Lincoln T.; Field, Loren J.; Pritchard, Catrin A.; Guerin, Christopher J.; Rubart, Michael; Soonpaa, Mark H.; Ang, Keng Leong; Galiñanes, Manuel.

In: PLoS One, Vol. 3, No. 4, e1929, 16.04.2008.

Research output: Contribution to journalArticle

Shenje, LT, Field, LJ, Pritchard, CA, Guerin, CJ, Rubart, M, Soonpaa, MH, Ang, KL & Galiñanes, M 2008, 'Lineage tracing of cardiac explant derived cells', PLoS One, vol. 3, no. 4, e1929. https://doi.org/10.1371/journal.pone.0001929
Shenje LT, Field LJ, Pritchard CA, Guerin CJ, Rubart M, Soonpaa MH et al. Lineage tracing of cardiac explant derived cells. PLoS One. 2008 Apr 16;3(4). e1929. https://doi.org/10.1371/journal.pone.0001929
Shenje, Lincoln T. ; Field, Loren J. ; Pritchard, Catrin A. ; Guerin, Christopher J. ; Rubart, Michael ; Soonpaa, Mark H. ; Ang, Keng Leong ; Galiñanes, Manuel. / Lineage tracing of cardiac explant derived cells. In: PLoS One. 2008 ; Vol. 3, No. 4.
@article{01062dc5628e406ea5c18bae7e92928b,
title = "Lineage tracing of cardiac explant derived cells",
abstract = "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 immunofluorescence is not adequate to identify the source, fate and function of adult cardiac explant derived cells.",
author = "Shenje, {Lincoln T.} and Field, {Loren J.} and Pritchard, {Catrin A.} and Guerin, {Christopher J.} and Michael Rubart and Soonpaa, {Mark H.} and Ang, {Keng Leong} and Manuel Gali{\~n}anes",
year = "2008",
month = "4",
day = "16",
doi = "10.1371/journal.pone.0001929",
language = "English (US)",
volume = "3",
journal = "PLoS One",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "4",

}

TY - JOUR

T1 - Lineage tracing of cardiac explant derived cells

AU - Shenje, Lincoln T.

AU - Field, Loren J.

AU - Pritchard, Catrin A.

AU - Guerin, Christopher J.

AU - Rubart, Michael

AU - Soonpaa, Mark H.

AU - Ang, Keng Leong

AU - Galiñanes, Manuel

PY - 2008/4/16

Y1 - 2008/4/16

N2 - 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 immunofluorescence is not adequate to identify the source, fate and function of adult cardiac explant derived cells.

AB - 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 immunofluorescence is not adequate to identify the source, fate and function of adult cardiac explant derived cells.

UR - http://www.scopus.com/inward/record.url?scp=44849088840&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=44849088840&partnerID=8YFLogxK

U2 - 10.1371/journal.pone.0001929

DO - 10.1371/journal.pone.0001929

M3 - Article

C2 - 18414652

AN - SCOPUS:44849088840

VL - 3

JO - PLoS One

JF - PLoS One

SN - 1932-6203

IS - 4

M1 - e1929

ER -