成纤维细胞生长因子和胰岛素促小鼠软骨细胞增殖分化的效应

背景根据软骨为单一类型细胞组成的无血管组织,对氧和营养的供应要求较低,同种免疫性较低,体内功能较简单,易于在体外构建成可供移植的人工移植细胞系等特点,从建立用于组织工程的通用人类同种软骨细胞系着手,为人工组织和人工器官走向标准化和批量生产奠定基础.目的探讨碱性成纤维细胞生长因子(basicfibroblast growth factor,bFGF)和胰岛素在原代培养软骨细胞的增殖过程中的效应,为组织工程中细胞因子的应用与作用进行评估.设计以细胞为研究对象,分组对照重复观察测量.单位一所大学的组织移植与免疫实验室.材料实验于2002-11/2003-05在暨南大学组织移植与免疫教育部重点实验室完成,研究对象为随机获取的培养的软骨细胞.方法在低血清培养基培养条件下进行小鼠原代软骨细胞培养,采用WST1比色法和荧光染色法观察不同浓度bFGF和胰岛素对软骨细胞增殖的影响.主要观察指标主要结局①bFGF对小鼠原代软骨细胞增殖影响.②胰岛素对小鼠原代软骨细胞增殖的影响.次要结局软骨细胞形态学观察.结果原代软骨细胞在4g/L胎牛血清培养基培养条件下,不同浓度的bFGF和胰岛素对软骨细胞的增殖有剂量相关性.形态学观察表明,bFGF和胰岛素不仅促进细胞增殖,而且可使细胞分泌基质增多.结论胰岛素和bFGF均能促进软骨细胞的增殖.

Effect of basic fibroblast growth factor and insulin on the proliferation and differentiation of mouse chondrocytes

BACKGROUND: Based on the characteristics of cartilage tissue, such as consisting of single type of cells, the cartilage cells or chondrocyte, absence of blood vessel, rather low consumption level of oxygen and nutrition, low level of allo-immunocompetence and simple function in vivo, it seems to be easy for cartilage cell lines to be established for tissue and cell transplantation. We want to set up a cell line with the purpose of current use in tissue engineering in vitro. It will provide the basis for artificial tissue and organ that will become to be standardized and yielded in batch.OBJECTIVE: To explore the potential stimulatory effects of basic fibroblast growth factor(bFGF) and insulin on the proliferation and differentiation in primary culture mice chondrocytes in vitro. The effect and application of the cell factors will be evaluated for tissue engineering.DESIGN: A grouping controlled and repeated trial was conducted with the cells as the subjects.SETTING: Key laboratory of tissue transplantation and immunology of a college.MATERIAIS: The experiment was completed in the Key Laboratory of Tissue Transplantation and Immunology of the Ministry of Education, Jinan University from November 2002 to May 2003. Cultured cartilage cells at random were obtained as the study objects.METHODS: Mice cartilage cells were cultured in medium at the minimum concentrations of serum. The effects of different concentration of bFGF and insulin on the proliferation and differentiation in mice cartilage cells were observed with WST1 and immunofluorescence staining.MAIN OUTCOME MEASURES: Primary results: ① Effect of bFGF on proliferation of primary cultured mice cartilage cells. ② Effect of insulin on proliferation of primary cultured mice cartilage cells. Secondary results:morphological observation of cartilage cells RESULTS: Primary cultured mice cartilage cells were cultured in medium at the minimum concentration of serum(4 g/L fatal bovine serum). It was found that bFGF and insulin might play an important role on the proliferation and growth of mice cartilage cells in a dose-dependent manner. In addition, morphological observation of cartilage cells showed that both bFGF and insulin not only promoted the proliferation of the cells but also enhanced the matrix secretion of cartilage cells.CONCLUSION: Both bFGF and insulin can stimulate the proliferation of cartilage cells in vitro.