title: [Study of 5-bromodeoxyuridine labeling of endothelial progenitor cells from the circulating blood from tooth movement rat].
abstract: 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.
authors.affiliation: Dept. of Orthodontics, School of Stomatology, Jilin University, Changchun 130041, China.
authors.forename: Chao
authors.initials: C
authors.lastname: Liu
authors.forename: Lei-Ying
authors.initials: LY
authors.lastname: Miao
authors.forename: Xin-Hua
authors.initials: XH
authors.lastname: Sun
authors.forename: Jin-Zhong
authors.initials: JZ
authors.lastname: Liu
mesh_terms: 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
chemical_list: D001973:Bromodeoxyuridine