title: Whole bone marrow cell culture: A convenient protocol for thein vitroexpansion of endothelial progenitor cells.
abstract: The number and function of endothelial progenitor cells (EPCs) may be a predictive factor for the severity and outcome of cardiovascular disease. However, the manipulation of bone marrow mononuclear cell (BMMC) cultures for EPCs is an elaborate and difficult procedure in small experimental animals. The present study aimed to assess the feasibility of whole bone marrow cell (WBMC) culture for expanding EPCs in small experimental animals. C57BL/6 mice (age, 3-4 weeks; weight, 9.47±0.76 g) were used as the experimental animals, and WBMCs were isolated from the femora and tibiae and cultured in endothelial cell growth medium-2. A BMMC culture for EPCs was used as a control. EPC growth, phenotype and functions were assessedin vitroandin vivo. The results demonstrated that EPCs were easily obtained from a WBMC culturein vitro. The cells exhibited similar growth and biological characteristics when compared with the EPCs derived from the traditional BMMC culture system. Thus, the cells were able to simultaneously bind to lectin and cause phagocytosis of acetylated-low density lipoproteins. In addition, the cells exhibited high expression levels of cluster of differentiation 34 and fetal liver kinase 1, and possessed similar functional properties to BMMC-derived EPCs, including vascular network formation, proliferation, adhesion and migration abilitiesin vitro. Thus, WBMC-derived EPCs can improve the outcome of pulmonary vascular disease when transplanted into a monocrotaline-induced pulmonary hypertension mouse model. The results of the present study indicated that the WBMC culture system is a more convenient and effective method of obtaining and expanding EPCs compared with BMMC culture, with the advantage of a simplified procedure.
authors.affiliation: Laboratory of Tissue Engineering and Stem Cells, Guiyang Medical College, Guiyang, Guizhou 550004, P.R. China ; Department of Pediatrics, The General Hospital of Huabei Oil Field Company, Renqiu, Hebei 062552, P.R. China.
authors.forename: Jun-Feng
authors.initials: JF
authors.lastname: Liu
authors.affiliation: Department of Cardiology, Beijing Children's Hospital, Capital Medical University, Beijing 100045, P.R. China.
authors.forename: Zhong-Dong
authors.initials: ZD
authors.lastname: DU
authors.affiliation: Department of Cardiology, Beijing Children's Hospital, Capital Medical University, Beijing 100045, P.R. China.
authors.forename: Zhi
authors.initials: Z
authors.lastname: Chen
authors.affiliation: Laboratory of Tissue Engineering and Stem Cells, Guiyang Medical College, Guiyang, Guizhou 550004, P.R. China.
authors.forename: Zhi-Xu
authors.initials: ZX
authors.lastname: He
keywords: bone marrow; endothelial progenitor cells; model; mouse