[Study of 5-bromodeoxyuridine labeling of endothelial progenitor cells from the circulating blood from tooth movement rat].
OBJECTIVE
To investigate the optimal dosage and timing for 5-bromodeoxyuridine (BrdU) labeling of endothelial progenitor cells (EPCs) from rat circulating blood.


METHODS
The animal model for rat tooth movement was established. EPCs were obtained by density gradient centrifugation. The expressions of specific antigens on cell surface were analysed by immunocytochemistry and fluorescenceochemistry. EPCs were incubated with BrdU at different concentrations (5, 10, 15 micromol/L) for different incubating time (12, 24, 48, 72, 96 h) to identify the optimal BrdU concentration and incubating time for cell labeling. Immunohistochemistry was performed to calculate the labeling index (LI).


RESULTS
The culture cell positively expressed CD34, CD133 and could be shown to endocytose DiI-ac-LDL, FITC-UEA-1. Incubation of the EPCs with BrdU at 10 micromol/L and for an optimal length of 72 h appeared to achieve the highest LI (66.8+/-2.9)%, which was significantly higher than group of 5 micromol/L (P<0.05), while there was no significant difference between the group of 15 micromol/L and 10 micromol/L (P>0.05).


CONCLUSIONS
EPCs can be isolated from tooth movement rat circulating blood and cultured. Incubation of the EPCs with BrdU at 10 micromol/L and for an optimal length of 72 h appeared to achieve the optimal LI. This provides a foundation for us to investigate the mechanism of chemiotaxis and differentiation for EPCs.
Liu
Chao
C
Dept. of Orthodontics, School of Stomatology, Jilin University, Changchun 130041, China.
Miao
Lei-Ying
LY
Sun
Xin-Hua
XH
Liu
Jin-Zhong
JZ
D000818:Animals; D001973:Bromodeoxyuridine; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002478:Cells, Cultured; D042783:Endothelial Cells; D066026:Endothelial Progenitor Cells; D051381:Rats; D013234:Stem Cells; D014087:Tooth Movement Techniques
D001973:Bromodeoxyuridine