Isolation, characterization, and transplantation of cardiac endothelial cells

Busadee Pratumvinit, Kanit Reesukumal, Kajohnkiart Janebodin, Nicholas Ieronimakis, Morayma Reyes Gil

Research output: Contribution to journalArticle

5 Citations (Scopus)

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.

Original languageEnglish (US)
Article number359412
JournalBioMed Research International
Volume2013
DOIs
StatePublished - 2013
Externally publishedYes

Fingerprint

Endothelial cells
Heart Transplantation
Endothelial Cells
Flow cytometry
Microvessels
Flow Cytometry
Cytology
Intra-Arterial Injections
Cell Separation
Blood Vessels
Cell Biology
Digestion

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Cite this

Isolation, characterization, and transplantation of cardiac endothelial cells. / Pratumvinit, Busadee; Reesukumal, Kanit; Janebodin, Kajohnkiart; Ieronimakis, Nicholas; Reyes Gil, Morayma.

In: BioMed Research International, Vol. 2013, 359412, 2013.

Research output: Contribution to journalArticle

Pratumvinit, Busadee ; Reesukumal, Kanit ; Janebodin, Kajohnkiart ; Ieronimakis, Nicholas ; Reyes Gil, Morayma. / Isolation, characterization, and transplantation of cardiac endothelial cells. In: BioMed Research International. 2013 ; Vol. 2013.
@article{1e63c70831ce4b7e871cd1dbcf06a096,
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.",
author = "Busadee Pratumvinit and Kanit Reesukumal and Kajohnkiart Janebodin and Nicholas Ieronimakis and {Reyes Gil}, Morayma",
year = "2013",
doi = "10.1155/2013/359412",
language = "English (US)",
volume = "2013",
journal = "BioMed Research International",
issn = "2314-6133",
publisher = "Hindawi Publishing Corporation",

}

TY - JOUR

T1 - Isolation, characterization, and transplantation of cardiac endothelial cells

AU - Pratumvinit, Busadee

AU - Reesukumal, Kanit

AU - Janebodin, Kajohnkiart

AU - Ieronimakis, Nicholas

AU - Reyes Gil, Morayma

PY - 2013

Y1 - 2013

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=84889012411&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84889012411&partnerID=8YFLogxK

U2 - 10.1155/2013/359412

DO - 10.1155/2013/359412

M3 - Article

VL - 2013

JO - BioMed Research International

JF - BioMed Research International

SN - 2314-6133

M1 - 359412

ER -