碱性成纤维细胞因子对猪弹性软骨细胞增殖和成软骨能力的影响  

The effect of basic fibroblast growth factor on proliferation and chondrogenetic potency of porcine elastic chondrocytes in vitro

作　　者：丁小邦[1] 夏万尧[1] 陈兵[1] 崔磊[1] 刘伟 商庆新[1] 曹谊林[1] 

机构地区：[1]第二医科大学附属第九人民医院整复外科上海市组织工程重点实验室,上海200011

出　　处：《中华实验外科杂志》2002年第6期573-574,I004,共3页Chinese Journal of Experimental Surgery

基　　金：国家重点基础研究发展规划资助项目 (G19990 54 30 4 )

摘　　要：目的　研究碱性成纤维细胞因子 (b FGF)对体外培养的弹性软骨细胞增殖和成软骨能力的影响 ,探讨b FGF对软骨组织工程的意义。方法　细胞取自猪耳软骨 ,用含 1 0、2 5、50 μg/L3种浓度b FGF的培养液 ,体外培养至第 3代 ,从细胞形态、数量和基质分泌等方面进行研究。结果　含b FGF组细胞贴壁呈类成纤维细胞样 ,撤去b FGF ,细胞形态可恢复正常 ;含b FGF各组在第 3代 ,细胞总数是无b FGF组的 60～ 70倍 (P <0 .0 1 ) ,但 3种不同浓度b FGF对细胞增殖差异无显著性 ;培养液氨基糖氨多糖 (GAG)含量测定各组差异无显著性 ;免疫组织化学和原位杂交示各组第 3代软骨细胞仍有Ⅱ型胶原表达。结论　应用b FGF能在 3周内从少量软骨组织获取大量软骨细胞 ,且未改变细胞表型和功能 ,对软骨组织工程有重要意义 ;在 1 0、2 5、50 μg/L 3种浓度中 ,1 0μg/L为bObjective To study the effect of basic fibroblast growth factor(b FGF) on proliferation and chondrogenetic potency of porcine elastic chondrocytes in vitro.Methods Cells were harvested from the elastic auricular cartilage of swines.Viability and quantification of the cells were determined.Cells were plated at equal concentration in different plates.The effect of b FGF on cell growth and chondrogenetic potency was studied in the following groups:Group 1(control group) receiving Ham's F 12 with supplements but no b FGF;Group 2 receiving 10?μg/L b FGF;Group 3 receiving 25?μg/L b FGF;Group 4 receiving 50?μg/L b FGF.Results Chondrocytes displayed a morphology similar to fibroblasts in the media containing b FGF.After 3 day culture these cells returned to the original shape;Compared to the group 1,all of the other groups receiving b FGF approximately had a 70 fold increase in total cell number at the third passage;The cells in the third passage were still able to produce type Ⅱ collagen demonstrated by immunohistochemistry and in situ hybridization;The quantity of GAG secreted by cells in all 4 groups had no difference.Conclusions Chondrocytes can be vastly expanded from a small number of cells without losing their phenotype when b FGF was added,indicating that b FGF offers the possibility to provide large amounts of seed cells which can be expanded from small amount of harvested tissue.Among the three different concentrations of b FGF,the optimal concentration of b FGF is 10?μg/L.

关 键 词：碱性成纤维细胞因子 猪 弹性软骨细胞 成软骨能力 组织工程 细胞增殖 

分 类 号：R318[医药卫生—生物医学工程]