摘要
目的 研究不同代体外培养关节软骨细胞成软骨能力 ,选择软骨组织工程最佳种子细胞。方法 将兔膝关节软骨消化分离出软骨细胞 ,传代培养至第 6代 ,每代细胞与模板支架材料Pluron icF 12 7混匀形成复合物 ,使细胞浓度达 5× 10 7/ml,将复合物注射于自体兔背部皮下 ,9周后取材 ,进行大体观察、新生软骨湿重与体积测量、组织学观察。结果 体外培养关节软骨细胞随传代次数的增加 ,形态从圆形逐渐向梭形转变。 1～ 3代软骨细胞成软骨能力最强 ,新生软骨的湿重与体积最大 ,组织结构与正常软骨相同 ;第 4代成软骨能力减弱 ,软骨基质分泌减少 ,新生软骨湿重和体积减小 ,与前 3代比较差异有显著性 ;第 5代仅形成分散于纤维组织内的软骨细胞团 ;第 6代则未发现有软骨样组织形成。结论 ①在体外单层培养条件下 ,随传代次数的增加关节软骨细胞逐渐去分化 ,成软骨能力逐渐降低直到完全丧失。②前 3代软骨细胞均具备形成正常软骨的能力 ,都可用作组织工程的种子细胞 ,但传代培养的目的是使细胞得以最大限度扩增 ,因此第
Objective To research the chondrogenic ability of chondrocytes cultured in vitro of different generations and to define the optimal generation of chondrocytes for tissue engineering.Method The chondrocytes isolated from cartilage in rabbit's knee joint were cultured and propagated from 1st to 6th generations in vitro.The chondrocyte templete composite which consisted of each generation chondrocytes and Pluronic F 127 surpensions with the cell concentration of 5×10 7/ml were subcutanously injected into the dorsum of autologous rabbits.After nine weeks,the histologic construture of neocartilage had been observed and its the wet weight and volume had been measured that were formed respectively by every generational chondrocytes.Result With the increasing of generations,the morphology of chondrocytes was changing gradually in the condition of mono layer culture in vitro.The chondrogenic ability of 1～3 generations chondrocytes was the most strong and the wet weight and volume of the neocartilage was the biggest,and their histologic constructure was same as the natural cartilage.The chondrogenic ability of 4th chondrocytes decreased and chondral matrix secreated by 4th generation chondrocytes was less much than that of 1～3th generations,so was the wet weight and volume of 4th generation.There were significant differentiation between 1～3th generations and the 4th generation.Only a little of neochartilage was formed in 5th generation,but no neochartilage formed in 6th generations.Conclusion ①In the condition of mono layer culture in vitro,the chondrocytes show the trend of differentiation and the chondrogenic ability is decreasing generation by generation.②The 3th generation chondrocytes are the optimal seed cell for cartilage tissue engineering.
出处
《潍坊医学院学报》
2001年第4期260-262,W005,共4页
Acta Academiae Medicinae Weifang
基金
山东省卫生厅资助课题 (课题号 :1 999A2 5)
关键词
关节
软骨细胞
体外培养
种子细胞
Articulation
Seed cell
Chondrocyte
In vitro culture
