纤维蛋白凝胶复合碱性成纤维细胞生长因子与大鼠胎肢细胞碱性磷酸酶的活性

背景:纤维蛋白是一种天然的可生物降解、组织相容性好的高分子材料.碱性成纤维细胞生长因子是一种对中胚层和神经外胚层细胞促分裂作用的多肽生长因子.两者结合有利于干细胞在高度交联的凝胶支架内迁移,并且促进其增殖与分化.目的:观察纤维蛋白凝胶复合碱性成纤维细胞生长因子对大鼠胎肢细胞碱性磷酸酶活性的影响.方法:实验分为4组:纤维蛋白凝胶组,纤维蛋白凝胶中加入正常培养基.纤维蛋白凝胶复合碱性成纤维细胞生长因子组,在纤维蛋白凝胶中加入含10 μg/L碱性成纤维细胞生长因子培养基.碱性成纤维细胞生长因子组,加入10 μg/L 碱性成纤维细胞生长因子培养基.对照组,采用无凝胶正常培养基培养.无菌条件下分离培养大鼠胎肢细胞,取第3代细胞接种于以上培养基中.分别在第3,5,7,14,21,28天检测碱性磷酸酶活性、mRNA表达及细胞形态观察.结果与结论:在有纤维蛋白凝胶组内培养至7 d细胞具有较长突起且连接成网,而在无纤维蛋白凝胶组内的细胞呈纺锤形或立方形;纤维蛋白凝胶复合碱性成纤维细胞生长因子培养7,14,21 d碱性磷酸酶活性高于单纯纤维蛋白凝胶组和单纯碱性成纤维细胞生长因子培养组(P < 0.05).各时间点纤维蛋白凝胶复合碱性成纤维细胞生长因子组碱性磷酸酶mRNA表达均较对照组增强(P < 0.01).提示纤维蛋白凝胶复合碱性成纤维细胞生长因子有利于细胞形态发生,并明显增强碱性磷酸酶活性.

Effects of fibrin gel combined with basic fibroblast growth factor on alkaline phosphatase activity in fetal rat limb cells

BACKGROUND: Fibrin is a natural biodegradable polymer material with good histocompatibility. Basic fibroblast growth factor (bFGF) is an important polypeptide growth factor with mitogenic effect. Fibrin gel (FG) combined with bFGF is conducive to cell migration and promotes proliferation and differentiation. OBJECTIVE: To investigate the effects of fibrin gel combined with bFGF on alkaline phosphatase activity (ALP) in fetal rat limb cells. METHODS: Four groups were included: FG group: fetal rat limb cells were seeded in FG (three-dimensional, 3D) with normal medium; FG + bFGF group: cells were seeded in FG containing 10 μg/L bFGF for 24-hour 3D culture; bFGF group: cells were seeded in 10 μg/L bFGF medium; normal control group: cells were seeded in normal culture medium. Under aseptic condition, rat fetal limb cells were isolated. Passage 3 cells were used for each group above mentioned. ALP activity, mRNA expression and cell morphology were analyzed at 3, 5, 7, 14, 21, and 28 days after culture. RESULTS AND CONCLUSION: The cells in the FG had more processes and formed an interconnected network after 7 days of incubation, while the cells in no gels remained cuboidal or "spindle-shaped". At 7, 14, and 21 days, ALP activity was greater in the FG+ bFGF group than in the other groups (P < 0.05). At each time point, mRNA expression of ALP was greater in the FG+ bFGF group than in the control group (P < 0.01). These findings indicate that FG combined with bFGF is conducive to cell morphogenesis and markedly enhances ALP activity.