Publication: Isolation, characterization, and transplantation of cardiac endothelial cells
Issued Date
2013-12-09
Resource Type
ISSN
23146141
23146133
23146133
Other identifier(s)
2-s2.0-84889012411
Rights
Mahidol University
Rights Holder(s)
SCOPUS
Bibliographic Citation
BioMed Research International. Vol.2013, (2013)
Suggested Citation
Busadee Pratumvinit, Kanit Reesukumal, Kajohnkiart Janebodin, Nicholas Ieronimakis, Morayma Reyes Isolation, characterization, and transplantation of cardiac endothelial cells. BioMed Research International. Vol.2013, (2013). doi:10.1155/2013/359412 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/31141
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Title
Isolation, characterization, and transplantation of cardiac endothelial cells
Abstract
Isolation and ex vivo expansion of cardiac endothelial cells have been a recurrent challenge due to difficulties in isolation, cell heterogeneity, lack of specific markers to identify myocardial endothelial cells, and inadequate conditions to maintain long-term cultures. Herein, we developed a method for isolation, characterization, and expansion of cardiac endothelial cells applicable to study endothelial cell biology and clinical applications such as neoangiogenesis. First, we dissociated the cells from murine heart by mechanical disaggregation and enzymatic digestion. Then, we used flow cytometry coupled with specific markers to isolate endothelial cells from murine hearts. CD45+ cells were gated out to eliminate the hematopoietic cells. CD31+/Sca-1+ cells were isolated as endothelial cells. Cells isolated from atrium grew faster than those from ventricle. Cardiac endothelial cells maintain endothelial cell function such as vascular tube formation and acetylated-LDL uptake in vitro. Finally, cardiac endothelial cells formed microvessels in dorsal matrigel plug and engrafted in cardiac microvessels following intravenous and intra-arterial injections. In conclusion, our multicolor flow cytometry method is an effective method to analyze and purify endothelial cells from murine heart, which in turn can be ex vivo expanded to study the biology of endothelial cells or for clinical applications such as therapeutic angiogenesis. © 2013 Busadee Pratumvinit et al.