[Sorting of lymphatic endothelial progenitor cells from canine peripheral blood and their differentiation induction towards endothelial cells].
OBJECTIVE
To investigate the biological properties of CD34+/CD133 +/VEGFR-3 + lymphatic endothelial progenitor cells in peripheral blood and explore the effects of the VEGF-C/VEGFR-3 signaling pathway on differentiation of lymphatic endothelial progenitor cells to lymphatic endothelial cells.


METHODS
Mononuclear cells were isolated from peripheral blood by density centrifugation with Percoll solution, and VEGFR-3+ cells were sorted from them with flow cytometry. Differentiation of VEGFR-3+ cells was induced with VEGF-C. The morphology and ultrastructures of the cells were observed with scanning and transmission electron microscopes. Expression of surface markers were examined with a confocal laser scanning microscope.


RESULTS
VEGFR-3+ cells expressed CD34 and CD133 antigen. The percentage of CD34+/ VEGFR-3+ and VEGFR-3+/CD133+ cells were 0.13% and 0.08% of peripheral blood MNC respectively. The size of CD34+/CD133+/VEGFR-3+ cells was about 15 microm in the diameter. After induction with VEGFC, they were increased. The cells were shuttle-like in shape and extended the lamellipodia and many filopodia. After 1 week induction with VEGF-C, they expressed coagulation factor VII related antigen, and at 2 week induction, they showed caveolae on the surface and Weibel-Palade body inside the cells. The specific lymphatic endothelial marker LYVE-1 was expressed on the cells, and no longer expressed CD133.


CONCLUSIONS
CD34+/CD133+/VEGFR-3+ lymphatic endothelial progenitor cells from peripheral blood may differentiate into lymphatic endothelial cells. The VEGF-C/VEGFR-3 signaling pathway has important effects on the differentiation of the lymphatic endothelial progenitor cells.
Liu
Rui
R
Sorting of lymphatic endothelial progenitor cells from canine peripheral blood and their differentiation induction towards endothelial cells
Tan
Yu-Zhen
YZ
Wang
Hai-Jie
HJ
Zhang
Meihua
M
D000071916:AC133 Antigen; D000818:Animals; D015703:Antigens, CD; D018952:Antigens, CD34; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D004285:Dogs; D042783:Endothelial Cells / Q000166:cytology* / Q000378:metabolism; D005434:Flow Cytometry; D006023:Glycoproteins; D042601:Lymphatic Vessels / Q000166:cytology*; D008297:Male; D010455:Peptides; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D042582:Vascular Endothelial Growth Factor C / Q000378:metabolism / Q000494:pharmacology; D040321:Vascular Endothelial Growth Factor Receptor-3 / Q000378:metabolism
D000071916:AC133 Antigen; D015703:Antigens, CD; D018952:Antigens, CD34; D006023:Glycoproteins; D010455:Peptides; D042582:Vascular Endothelial Growth Factor C; D040321:Vascular Endothelial Growth Factor Receptor-3