Combination of rat tail collagen and platelet-derived growth factor-BB for culture of human umbilical vein endothelial cells / 中国组织工程研究

ABSTRACT

BACKGROUND:Rat tail colagen type I can promote the increase of muscle fiber cels and migration and tube formation of endothelial cels, which is speculated to provide a more suitable internal environment for the growth of cels. OBJECTIVE:To observe the effect and safety of platelet-derived growth factor-BB (PDGF-BB) with rat rail colagen against apoptosis of human umbilical vein endothelial celsin vitro. METHODS:The passage 4 human umbilical vein endothelial cels were cultured in the medium of rat rail colagen and the reduction ratio in different time points was detected by Alamar Blue. The passage 4 human umbilical vein endothelial cels were divided into four groups and cultured in 24-wel culture plates control group, PDGF-BB group, H2O2 group, PDGF-BB+colagen group. H2O2 was used to induce cel apoptosis in al the groups. Western blot was used to detect the expression of PDGF-BB, apoptosis-related protein and anti-apoptosis protein after 72 hours. Meanwhile, TUNEL method was used to detect cel apoptotic rate. RESULTS AND CONCLUSION:The tube formation in the human umbilical vein endothelial cels was more than that cultured in normal medium (P < 0.05). Cels cultured with rat tail colagen showed similar growth to normal control cels, indicating rat tail colagen had no cytotoxicity. The expressions of PDGF-BB, Bcl-2, and p-Akt in the PDGF-BB+colagen group were significantly higher than those in the other three groups (P < 0.05), but the expression of Bax was lower than that in the other three groups (P < 0.05). The apoptotic rate in the PDGF-BB+colagen group was lower than the PDGF-BB group and H2O2 group (P < 0.05). These findings indicate that rat tail colagen has no cytotoxicity to human umbilical vein endothelial cells, and rat tail collagen combined with PDGF-BB can predominantly enhance anti-apoptosis effects.