Isolation and transplantation of autologous circulating endothelial cells into denuded vessels and prosthetic grafts: implications for cell-based vascular therapy.
BACKGROUND
Blood-borne endothelial cells originating from adult bone marrow were reported previously. These cells have the properties of an endothelial progenitor cell (EPC) and can be mobilized by cytokines and recruited to sites of neovascularization, where they differentiate into mature endothelial cells. Current protocols for isolation of EPCs from peripheral blood rely on enrichment and selection of CD34+ mononuclear cells.


RESULTS
In this report, we describe a streamlined method for the isolation and expansion of EPCs from peripheral blood and evaluate their therapeutic potential for autologous cell-based therapy of injured blood vessels and prosthetic grafts. A subset of unfractionated mononuclear cells exhibited the potential to differentiate in vitro into endothelial cells under selective growth conditions. The cells were efficiently transduced ex vivo by a retroviral vector expressing the LacZ reporter gene and could be expanded to yield sufficient numbers for therapeutic applications. Transplantation of these cells into balloon-injured carotid arteries and into bioprosthetic grafts in rabbits led to rapid endothelialization of the denuded vessels and graft segments, resulting in significant reduction in neointima deposition.


CONCLUSIONS
We conclude that transplantation of EPCs may play a crucial role in reestablishing endothelial integrity in injured vessels, thereby inhibiting neointimal hyperplasia. These findings may have implications for novel and practical cell-based therapies for vascular disease.
Department of Medicine, Brigham and Women's Hospital and Harvard Medical School, 75 Francis St, Boston, Mass 02115, USA.
Daniel P
DP
Griese
Afshin
A
Ehsan
Luis G
LG
Melo
Deling
D
Kong
Lunan
L
Zhang
Michael J
MJ
Mann
Richard E
RE
Pratt
Richard C
RC
Mulligan
Victor J
VJ
Dzau
D000800:Angioplasty, Balloon / Q000009:adverse effects; D000818:Animals; D018952:Antigens, CD34 / Q000096:biosynthesis; D001705:Bioprosthesis; D001808:Blood Vessels / Q000293:injuries* / Q000473:pathology; D016893:Carotid Stenosis / Q000209:etiology / Q000473:pathology / Q000517:prevention & control*; D002454:Cell Differentiation; D002455:Cell Division; D002470:Cell Survival; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000166:cytology / Q000637:transplantation*; D017930:Genes, Reporter; D006083:Graft Occlusion, Vascular / Q000209:etiology / Q000473:pathology / Q000517:prevention & control*; D006085:Graft Survival; D006965:Hyperplasia / Q000517:prevention & control; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000378:metabolism; D008297:Male; D011817:Rabbits; D012190:Retroviridae / Q000235:genetics; D014182:Transplantation, Autologous; D017539:Tunica Intima / Q000166:cytology / Q000473:pathology*; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics
D018952:Antigens, CD34; D001616:beta-Galactosidase