应用碱性成纤维细胞生长因子体外扩增猪耳软骨细胞老化的规律  

Rule of swine chondrocyte aging by using bFGF in vitro culture

作　　者：丁小邦[1] 高建华[1] 夏万尧[2] 陈兵[2] 周广东[2] 崔磊[2] 刘伟[2] 曹谊林[2] 

机构地区：[1]南方医科大学附属南方医院,广州510515 [2]上海二医大附属第九人民医院整复外科

出　　处：《中华实验外科杂志》2007年第3期321-323,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（G19990543000）

摘　　要：目的探讨碱性成纤维细胞生长因子（bFGF）体外扩增软骨细胞的老化规律。方法细胞取自猪耳弹性软骨，实验分两组：培养液含10ug/L b-FGF组（简称bFGF组），培养液无bF-GF组（简称对照组），分别收集两组多代细胞，按5×10^7个／ml浓度与30％Pluronic F-127／Ham’s F-12混合，按每一样本0．5ml注射到猪自体皮下，8周后取材，从大体观察、称重及组织学染色，对再生的软骨组织进行评价。结果bFGF组和对照组从第3代开始均未能形成软骨组织，bFGF组第2代软骨细胞和原代比较，在两周内扩增70倍，是对照组的12．7倍，且体内能形成很好的软骨。结论应用bFGF体外扩增的软骨细胞在第3代开始老化，失去成软骨能力，种子细胞应取第2代，可兼顾适量细胞数与具备成软骨能力。＂Objective To study the rule of swine chondrocyte aging by using bFGF in vitro culture. Methods Ceils were harvested from the elastic auricular cartilage of swines and divided into two groups：bFGF group with Ham＇s F-12 containing 10 ug/L b-FGF,and control group with Ham＇s F-12 not containing b-FGF. 5 × 10^7 cells/mL in different passages were mixed with 30% Pluronic F-127/Ham＇ s F- 12 ,and injected subcutaneously at 0. 5 mL per implant. Implants were harvested after 8 weeks of in vivo culture and then examined with histologic stains. Results Chondrocytes from passage 3 didn＇ t form cartilage in two groups. The number of cells doublings from passage 2 （ after 12.75 ± 1.26 days） in in vitro culture in the bFGF group and control group was 70 and 5.5 respectively. Using bFGF in vitro culture, cells were proliferated by 70 times after 2 weeks. Histologically, all the engineered cartilage was similar to the native elastic cartilage, Conclusion Chondroeytes from passage 3 using bFGF in vitro culture become aging. The largest quantity of seed cells for tissue-engineered cartilage is harvested from passage 2 using b- FGF in vitro culture.

关 键 词：碱性成纤维细胞生长因子 软骨细胞 老化 

分 类 号：R686[医药卫生—骨科学]