title: Comparisons of rabbit bone marrow mesenchymal stem cell isolation and culture methods in vitro.
abstract: Bone marrow mesenchymal stem cells (BMSCs) have great potential in tissue engineering and clinical therapy, and various methods for isolation and cultivation of BMSCs have been reported. However, the best techniques are still uncertain. Therefore, we sought the most suitable among the four most common methods for BMSC separation from rabbits. BMSCs were obtained from untreated whole bone marrow (BM) adherent cultures, 3 volumes of red blood cells (RBC) lysed with ammonium chloride, 6 volumes of RBC lysed with ammonium chloride, and Ficoll density gradient centrifugation. Then, isolated BMSCs were evaluated with respect to primary cell yield, number of CFU-F colonies, proliferative capacity, cell phenotype, and chondrogenic differentiation potential. Our data show that BMSCs were successfully isolated by all four methods, and each method was similar with regard to cell morphology, phenotype, and differentiation potential. However, BMSCs from untreated whole BM adherent cultures had greater primary cell yields, larger colonies, and the shortest primary culture time (P<0.05). Moreover, the 4(th) generation of cultured cells had the strongest proliferative activity, the fastest growth rate and the most numerous cells compared with other cell passage generations (P<0.05). In conclusion, untreated whole BM adherent cultures are best for rabbit BMSC isolation and the 4(th) generation of cells has the strongest proliferation capacity.
authors.affiliation: Department of Cardiothoracic Surgery, College of Clinical Medicine, Yangzhou University, Yangzhou, Jiangsu Province, China ; The Research Center for Translational Medicine, Yangzhou University, Yangzhou, Jiangsu Province, China.
authors.forename: Weidong
authors.initials: W
authors.lastname: Zhang
authors.affiliation: Department of Cardiothoracic Surgery, College of Clinical Medicine, Yangzhou University, Yangzhou, Jiangsu Province, China ; The Research Center for Translational Medicine, Yangzhou University, Yangzhou, Jiangsu Province, China.
authors.forename: Fangbiao
authors.initials: F
authors.lastname: Zhang
authors.affiliation: Department of Cardiothoracic Surgery, College of Clinical Medicine, Yangzhou University, Yangzhou, Jiangsu Province, China ; The Research Center for Translational Medicine, Yangzhou University, Yangzhou, Jiangsu Province, China.
authors.forename: Hongcan
authors.initials: H
authors.lastname: Shi
authors.affiliation: Department of Cardiothoracic Surgery, College of Clinical Medicine, Yangzhou University, Yangzhou, Jiangsu Province, China ; The Research Center for Translational Medicine, Yangzhou University, Yangzhou, Jiangsu Province, China.
authors.forename: Rongbang
authors.initials: R
authors.lastname: Tan
authors.affiliation: Department of Cardiothoracic Surgery, College of Clinical Medicine, Yangzhou University, Yangzhou, Jiangsu Province, China ; The Research Center for Translational Medicine, Yangzhou University, Yangzhou, Jiangsu Province, China.
authors.forename: Shi
authors.initials: S
authors.lastname: Han
authors.affiliation: Department of Cardiothoracic Surgery, College of Clinical Medicine, Yangzhou University, Yangzhou, Jiangsu Province, China ; The Research Center for Translational Medicine, Yangzhou University, Yangzhou, Jiangsu Province, China.
authors.forename: Gang
authors.initials: G
authors.lastname: Ye
authors.affiliation: Department of Cardiothoracic Surgery, College of Clinical Medicine, Yangzhou University, Yangzhou, Jiangsu Province, China ; The Research Center for Translational Medicine, Yangzhou University, Yangzhou, Jiangsu Province, China.
authors.forename: Shu
authors.initials: S
authors.lastname: Pan
authors.affiliation: Department of Cardiothoracic Surgery, College of Clinical Medicine, Yangzhou University, Yangzhou, Jiangsu Province, China ; The Research Center for Translational Medicine, Yangzhou University, Yangzhou, Jiangsu Province, China.
authors.forename: Fei
authors.initials: F
authors.lastname: Sun
authors.affiliation: Department of Cardiothoracic Surgery, College of Clinical Medicine, Yangzhou University, Yangzhou, Jiangsu Province, China ; The Research Center for Translational Medicine, Yangzhou University, Yangzhou, Jiangsu Province, China.
authors.forename: Xingchen
authors.initials: X
authors.lastname: Liu
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology*; D002448:Cell Adhesion; D018929:Cell Culture Techniques / Q000379:methods*; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods*; D002499:Centrifugation, Density Gradient; D016130:Immunophenotyping; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D011817:Rabbits