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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(basicfibroblastgrowthfactor,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
Background: According to the fact that cartilage is a kind of avascular tissue consisting of a single type of cell, it has lower requirement of supply of oxygen and nutrition, lower immunity of the same kind, simpler function in vivo and easy to be constructed in vitro into artificial transplanted cell line for transplantation Characteristics, from the establishment of universal human allogeneic cartilage cell line for tissue engineering to lay the foundation for the standardization and mass production of artificial tissues and artificial organs. OBJECTIVE: To investigate the effect of basic fibroblast growth factor (bFGF) and insulin on the proliferation of primary cultured chondrocytes, and to evaluate the role and application of cytokines in tissue engineering. Design: Cells were used as study objects, and the control group was repeated observation and measurement. Unit: a university transplant and immunology laboratory. MATERIALS: The experiment was performed at the Key Laboratory of Tissue Transplantation and Immunology of Ministry of Education, Jinan University from November 2002 to May 2003 with the aim of obtaining randomly cultured chondrocytes. Methods: Mouse primary chondrocytes were cultured in low serum medium. The effects of bFGF and insulin on proliferation of chondrocytes were observed by WST1 colorimetry and fluorescence staining. MAIN OUTCOME MEASURES: Main Outcomes: ① Effect of bFGF on Proliferation of Mouse Primary Chondrocytes. ② insulin on mouse primary chondrocyte proliferation. Secondary outcome: Chondrocyte morphological observation. Results: Primary chondrocytes cultured in 4g / L fetal bovine serum medium had different dosages of bFGF and insulin on the proliferation of chondrocytes in a dose-dependent manner. Morphological observations show that bFGF and insulin not only promote cell proliferation, but also make the cell secretion of matrix increased. Conclusion: Insulin and bFGF are both capable