Isolation and characterization of human coronary artery-derived endothelial cells in vivo from patients undergoing percutaneous coronary interventions.
OBJECTIVE
Human coronary artery-derived endothelial cells (ECs) seem to be the most appropriate cells for the pathogenesis study of coronary artery disease. But limited availability of endothelial tissue is a major constraint. In this study, we developed a method to isolate human coronary artery ECs in vivo from patients.


METHODS
Coronary guidewires were used to obtain EC samples from coronary arteries in 76 patients. Cells were eluted from wire tips and purified by immunomagnetic beads. Von Willebrand factor and CD31 were used as immunocytochemical markers to identify cells as endothelium. Cell viability was evaluated in terms of cell membrane integrity, energy-dependent uptake of DiI-labeled acetylated low-density lipoprotein, and apoptosis. Nitric oxide synthase (eNOS) expression and nitric oxide (NO) production of cells were detected to evaluate cell function.


RESULTS
About 96 coronary artery ECs were obtained per guidewire. Cells manifested endothelial morphology and immunoreactivity for von Willebrand factor and CD31 with good viability. But eNOS expression and NO production of cells were decreased.


CONCLUSIONS
Viable coronary endothelium could be obtained during routine percutaneous coronary interventions combined with immunomagnetic beads. These cells may be used for advanced cellular functional analyses such as immunocytochemistry and molecular biology. Such information could aid in understanding mechanisms of coronary artery diseases.
Yu
Shi-Yong
SY
Department of Cardiology, Xinqiao Hospital, Third Military Medical University, Chongqing, PR China.
Song
Yao-Ming
YM
Li
Ai-Min
AM
Yu
Xue-Jun
XJ
Zhao
Gang
G
Song
Ming-Bao
MB
Lin
Chun-Mei
CM
Tao
Chun-Rong
CR
Huang
Lan
L
D000368:Aged; D015906:Angioplasty, Balloon, Coronary; D017209:Apoptosis; D015415:Biomarkers / Q000378:metabolism; D051929:CD146 Antigen / Q000378:metabolism; D002470:Cell Survival; D002478:Cells, Cultured; D003331:Coronary Vessels / Q000166:cytology*; D042783:Endothelial Cells / Q000166:cytology* / Q000378:metabolism; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D018189:Immunomagnetic Separation / Q000379:methods*; D008297:Male; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism
D015415:Biomarkers; D051929:CD146 Antigen; C497902:MCAM protein, human; D019408:Platelet Endothelial Cell Adhesion Molecule-1; C496314:NOS3 protein, human; D052250:Nitric Oxide Synthase Type III