碱性成纤维细胞生长因子对无血清培养大鼠生长板软骨细胞的影响  

Effects of basic fibroblast growth factor on serum free cultured rat costochondral growth plate chondrocyte

作　　者：季煜华[1] 曾耀英[1] 俞瑜[1] 

机构地区：[1]暨南大学组织移植与免疫研究中心教育部重点实验室,广东广州510632

出　　处：《暨南大学学报（自然科学与医学版）》2005年第6期734-739,745,共7页Journal of Jinan University(Natural Science & Medicine Edition)

基　　金：973国家重点基础研究发展规划项目(G1999054303);广东省"十五"重大科技专项(A302020204)

摘　　要：目的:探讨碱性成纤维细胞生长因子(bFGF)用于无血清培养扩增软骨细胞的可行性及最适质量浓度.方法:分离、培养大鼠肋生长板软骨细胞(rat costochondral growth plate chondrocyte,RGC).细胞化学和免疫细胞化学染色的方法检测第1代RGC中蛋白多糖和Ⅱ型胶原的表达.观察0.1、1.0、10.0和100.0μg/L bFGF对无血清培养RGC形态的影响,并分别用3H-TdR和3H-Proline掺入法检测上述质量浓度bFGF对无血清培养RGC增殖和胶原合成的影响.结果:第1代RGC表达蛋白多糖和Ⅱ型胶原,保持了软骨细胞的分化表型.随着bFGF质量浓度的增加RGC的形态由多角形变为梭形和圆形.与无血清对照组相比,上述4个质量浓度的bFGF均显著促进RGC的增殖和胶原合成(P＜0.05),其中1.0和10.0μg/L bFGF促RGC增殖的作用与10%胎牛血清(FBS)对照组差异无显著性意义(P＞0.05),但促胶原合成作用较弱,相当于10%FBS对照组的60%左右(P＜0.05).结论:bFGF可用于无血清扩增软骨细胞,1.0～10.0 μg/L是适宜的质量浓度范围,但其促胶原合成作用较弱,还需添加其他因子以弥补促胶原合成作用的不足.Aim： To investigate the feasibility and find proper concentration of bFGF in expanding serum free cultured RGC（rat costochondral growth plate chondrocyte）. Methods： The RGCs were isolated and cultured from rat costochondral growth plate, The GAG（glycosaminoglycan） and collagen Ⅱ expression of the first passage RGC were detected by cytochemistry and immunocytochemistry （ICC）. The effects of 0. 1μg/L, 1.0μg/L, 10.0μg/L and 100.0μg/L bFGF on the morphology of the serum free cultured first passage RGC and the proliferation as well as collagen synthesis of the serum free cultured first passage RGC detected by ^3H- TdR,^3H- Proline incorporation were observed. Results： The first passage RGC expressed GAG and collagenⅡ, retained their phenotype in dvo. The morphology d RGC changes from polygon to spindle or round shape with the increasing concentration of bFGF. Compared with serum free control, the proliferation and collagen synthesis were markedly promoted by all the four above mentioned concentration bFGF（ P 〈 0.05） ; there was no difference between 1.0μg/L, 10.0μg/L bFGF and 10%FBS control in promoting RGC proliferation（ P 〉 0.05）, but their enhancement in collagen synthesis was weaker, the ^3H- Proline incorporation of 1.0μg/L, 10.0μg/L bFGF was about 60% as much as the 10%FBS control（P 〈0.05）. Conchlsion： bFGF is feasible for expanding serum free cultured RGC, the proper concentration ranges from 1.0μg/L to 10.0μg/L,but their effect on collagen synthesis is less than 10% FBS, other factors are needed to make up for this shortage.

关 键 词：肋生长板软骨细胞 碱性成纤维细胞生长因子 无血清培养 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]