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.
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U2 - 10.1371/journal.pone.0001929
DO - 10.1371/journal.pone.0001929
M3 - Article
C2 - 18414652
AN - SCOPUS:44849088840
SN - 1932-6203
VL - 3
JO - PloS one
JF - PloS one
IS - 4
M1 - e1929
ER -