OBJECTIVES: The aim of this study was to determine an optimal culture method for porcine bone marrow-derived endothelial progenitor cells (EPCs). MATERIALS AND METHODS: Mononuclear cells (MNCs) were isolated by density centrifugation and differentiated into EPCs in in vitro. At first-passage, EPCs were cultured at different cell densities (5 x 10(3), 1 x 10(4), 2 x 10(4) or 5 x 10(4)/cm(2)) and in basic medium (EGM, medium 199, DMEM or 1640) supplemented with FBS (2%, 5%, 10% or 20%) and different combinations of cytokines (VEGF, VEGF + bFGF, VEGF + bFGF + EGF, or VEGF + bFGF + EGF + IGF), the experiment being based on L(64) (4(21)) orthogonal design. RESULTS AND CONCLUSIONS: This demonstrated that the optimal culture method for our EPCs displayed higher expansion and migration rates as compared to other groups, by analysis of variance; that is, cultured at 1 x 10(4)/cm(2) in M199 supplemented with 10% FBS and VEGF + bFGF + IGF + EGF. Furthermore, percentage of positive cells stained by Dil-ac-LDL and FITC-UEA-1 was more than 65%, and as shown by immunohistochemistry, these cells also stained positively for CD133, CD34 and KDR. The present study indicates that the number and function of porcine EPCs significantly increased when using our optimized culture parameters.
OBJECTIVES: The aim of this study was to determine an optimal culture method for porcine bone marrow-derived endothelial progenitor cells (EPCs). MATERIALS AND METHODS: Mononuclear cells (MNCs) were isolated by density centrifugation and differentiated into EPCs in in vitro. At first-passage, EPCs were cultured at different cell densities (5 x 10(3), 1 x 10(4), 2 x 10(4) or 5 x 10(4)/cm(2)) and in basic medium (EGM, medium 199, DMEM or 1640) supplemented with FBS (2%, 5%, 10% or 20%) and different combinations of cytokines (VEGF, VEGF + bFGF, VEGF + bFGF + EGF, or VEGF + bFGF + EGF + IGF), the experiment being based on L(64) (4(21)) orthogonal design. RESULTS AND CONCLUSIONS: This demonstrated that the optimal culture method for our EPCs displayed higher expansion and migration rates as compared to other groups, by analysis of variance; that is, cultured at 1 x 10(4)/cm(2) in M199 supplemented with 10% FBS and VEGF + bFGF + IGF + EGF. Furthermore, percentage of positive cells stained by Dil-ac-LDL and FITC-UEA-1 was more than 65%, and as shown by immunohistochemistry, these cells also stained positively for CD133, CD34 and KDR. The present study indicates that the number and function of porcine EPCs significantly increased when using our optimized culture parameters.
Authors: Josef Neumüller; Sylvia-Emanuela Neumüller-Guber; Markus Lipovac; Wilhelm Mosgoeller; Monika Vetterlein; Margit Pavelka; Johannes Huber Journal: Histochem Cell Biol Date: 2006-06-10 Impact factor: 4.304
Authors: T Asahara; T Murohara; A Sullivan; M Silver; R van der Zee; T Li; B Witzenbichler; G Schatteman; J M Isner Journal: Science Date: 1997-02-14 Impact factor: 47.728
Authors: Konstantinos Stellos; Harald Langer; Karin Daub; Tanja Schoenberger; Alexandra Gauss; Tobias Geisler; Boris Bigalke; Iris Mueller; Michael Schumm; Iris Schaefer; Peter Seizer; Bjoern F Kraemer; Dorothea Siegel-Axel; Andreas E May; Stephan Lindemann; Meinrad Gawaz Journal: Circulation Date: 2007-12-17 Impact factor: 29.690
Authors: Tian Hang Luo; Yao Wang; Zheng Mao Lu; Hong Zhou; Xu Chao Xue; Jian Wei Bi; Li Ye Ma; Guo En Fang Journal: Crit Care Date: 2009-07-15 Impact factor: 9.097