title: A novel and feasible way to cultivate and purify endothelial progenitor cells from bone marrow of children with congenital heart diseases.
abstract: BACKGROUND
  Endothelial progenitor cells (EPCs) are used in vascular tissue engineering and clinic therapy. Some investigators get EPCs from the peripheral blood for clinic treatment, but the number of EPCs is seldom enough. We have developed the cultivation and purification of EPCs from the bone marrow of children with congenital heart disease, to provide enough seed cells for a small calibre vascular tissue engineering study.
  
  
  METHODS
  The 0.5-ml of bone marrow was separated from the sternum bone, and 5-ml of peripheral blood was collected from children with congenital heart diseases who had undergone open thoracic surgery. CD34+ and CD34+/VEGFR+ cells in the bone marrow and peripheral blood were quantified by flow cytometry. CD34+/VEGFR+ cells were defined as EPCs. Mononuclear cells in the bone marrow were isolated by Ficoll(®) density gradient centrifugation and cultured by the EndoCult Liquid Medium Kit(™). Colony forming endothelial cells was detected. Immunohistochemistry staining for Dil-ac-LDL and FITC-UEA-1 confirmed the endothelial lineage of these cells.
  
  
  RESULTS
  CD34+ and CD34+/VEGFR+ cells in peripheral blood were (0.07 ± 0.05)% and (0.05 ± 0.02)%, respectively. The number of CD34+ and CD34+/VEGFR+ cells in bone marrow were significantly higher than in blood, (4.41 ± 1.47)% and (0.98 ± 0.65)%, respectively (P < 0.0001). Many colony forming units formed in the culture. These cells also expressed high levels of Dil-ac-LDL and FITC-UEA-1.
  
  
  CONCLUSIONS
  This is a novel and feasible approach that can cultivate and purify EPCs from the bone marrow of children with congenital heart disease, and provide seed cells for small calibre vascular tissue engineering.
authors.affiliation: Department of Pediatric Cardiac Surgery, Beijing Anzhen Hospital, Capital Medical University, Beijing 100029, China.
authors.forename: Yong-Tao
authors.initials: YT
authors.lastname: Wu
authors.forename: Jing-Xing
authors.initials: JX
authors.lastname: Li
authors.forename: Shuo
authors.initials: S
authors.lastname: Liu
authors.forename: Yi
authors.initials: Y
authors.lastname: Xin
authors.forename: Zi-Jian
authors.initials: ZJ
authors.lastname: Wang
authors.forename: Jin
authors.initials: J
authors.lastname: Gao
authors.forename: Bing-Yang
authors.initials: BY
authors.lastname: Ji
authors.forename: Xiang-Ming
authors.initials: XM
authors.lastname: Fan
authors.forename: Qi-Wen
authors.initials: QW
authors.lastname: Zhou
mesh_terms: D000293:Adolescent; D000328:Adult; D018952:Antigens, CD34 / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology* / Q000378:metabolism; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D002648:Child; D002675:Child, Preschool; D042783:Endothelial Cells / Q000166:cytology* / Q000378:metabolism; D005260:Female; D005434:Flow Cytometry; D006330:Heart Defects, Congenital / Q000473:pathology*; D006801:Humans; D007150:Immunohistochemistry; D017493:Leukocyte Common Antigens / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D055815:Young Adult
chemical_list: D018952:Antigens, CD34; D017493:Leukocyte Common Antigens