Optimization of in vitro culture condition for porcine bone marrow-derived endothelial progenitor cells
10.3724/SP.J.1008.2010.00964
- Author:
Jian-Guo WU
1
Author Information
1. Department of General Surgery
- Publication Type:Journal Article
- Keywords:
Bone marrow;
Cell culture techniques;
Endothelial progenitor cells
- From:
Academic Journal of Second Military Medical University
2010;31(9):964-969
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To optimize the culture condition for porcine bone marrow-derived endothelial progenitor cells (EPCs), so as to lay a foundation for future study. Methods: Bone marrow was collected from porcine ilium (n = 5). Mononuclear cells (MNCs) were separated by density centrifugation and were induced to differentiate into EPCs in vitro. The influences of different cell inoculation densities (2 × 103/cm2, 5 × 103/cm2, 1 × 104/cm2, and 2 × 104/cm2), basic medium (EGM, medium 199, and DMEM), FBS (5%, 10%, 20%, and 30%), and combinations of cytokines (vascular endothelial growth factor [VEGF]+ basic fibroblast growth factor [bFGF], VEGF+stromal-derived factor [SDF], VEGF+bFGF+SDF, VEGF+bFGF+ insulin-like growth factor [IGF]+ epidermal growth factor [EGF], and VEGF+bFGF+SDF+IGF) on the proliferation and migration of EPCs were evaluated. EPCs were identified by morphology observation, fluorescent staining, and immunohistochemical method. Results: EPCs with the highest proliferation and migration ability were obtained under following condition: at a density of 1 × 104/cm 2 in M199 medium supplemented with 10% FBS and VEGF+bFGF+SDF+IGF. Furthermore, the percentage of double positive cells stained by Dil-ac-LDL and FITC-UEA-1 was higher than 76%, and these cells were also positively stained for CD133, CD34 and KDR immunohistochemically. Conclusion: Optimization of in vitro culture condition of porcine EPCs can increase the cell amount and improve their functions, paving a way for future related studies.
