CAJ | 학술논문

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(basic fibroblast 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4 g/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.
배경:근거연골위단일류형세포조성적무혈관조직,대양화영양적공응요구교저,동충면역성교저,체내공능교간단,역우재체외구건성가공이식적인공이식세포계등특점,종건립용우조직공정적통용인류동충연골세포계착수,위인공조직화인공기관주향표준화화비량생산전정기출.목적:탐토감성성섬유세포생장인자(basic fibroblast growth factor,bFGF)화이도소재원대배양연골세포적증식과정중적효응,위조직공정중세포인자적응용여작용진행평고.설계:이세포위연구대상,분조대조중복관찰측량.단위:일소대학적조직이식여면역실험실.재료:실험우2002-11/2003-05재기남대학조직이식여면역교육부중점실험실완성,연구대상위수궤획취적배양적연골세포.방법:재저혈청배양기배양조건하진행소서원대연골세포배양,채용WST1비색법화형광염색법관찰불동농도bFGF화이도소대연골세포증식적영향.주요관찰지표:주요결국:①bFGF대소서원대연골세포증식영향.②이도소대소서원대연골세포증식적영향.차요결국:연골세포형태학관찰.결과:원대연골세포재4 g/L태우혈청배양기배양조건하,불동농도적bFGF화이도소대연골세포적증식유제량상관성.형태학관찰표명,bFGF화이도소불부촉진세포증식,이차가사세포분비기질증다.결론:이도소화bFGF균능촉진연골세포적증식.
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.