以ITS为主要成分的无血清培养基对软骨细胞增殖和表型的影响  

Effects of Serum-free Culture Medium Containing ITS on Chondrocytes Proliferation and Phenotype

作　　者：刘瑾春[1,2] 康宁 肖苒 刘霞 曹谊林 

机构地区：[1]中国医学科学院整形外科医院研究中心,北京市100144 [2]新乡医学院河南省医用组织再生重点实验室,河南省新乡市453003

出　　处：《组织工程与重建外科杂志》2012年第3期121-125,共5页Journal of Tissue Engineering and Reconstructive Surgery

基　　金：国家自然科学基金青年科学基金(30801192);北京市自然科学基金(7102134);高等学校博士学科点专项科研基金(新教师基金课题;200800231078);卫生部临床学科重点项目;北京市科委重大计划项目(D090800046609003)

摘　　要：目的观察以ITS为主要成分的无血清培养基对软骨细胞增殖和表型的影响。方法培养猪耳软骨细胞,分为无血清组和含血清组,无血清组应用包含胰岛素铁硒传递蛋白(ITS)、地塞米松及维生素C为主要成分的无血清培养基;含血清组应用包含10%血清的常规培养基。观察平面和三维培养状态下无血清培养基对软骨细胞增殖、基质分泌和软骨表型维持的作用。结果平面培养条件下,无血清组的细胞增殖率出现下调,细胞表型不能维持。在三维培养情况下无血清组可以保持软骨细胞存活和软骨细胞的表型,Ⅱ型胶原和蛋白多糖组织学染色结果与含血清组无明显差异,软骨特异基因ColⅡ、Aggrecan的表达与含血清组无明显差异,并能够形成较好的软骨样组织。结论以ITS、地塞米松及维生素C为主要成分的无血清培养基,在三维培养条件下能基本保持软骨细胞的存活和表型,可用于软骨细胞的体外三维培养。Objective To investigate the effects of serum-free culture medium containing Insulin-Transferrin-Selenium on chondrocytes proliferation and phenotype. Methods P2 Chondrocytes were cultured in monolayer or in 3-D pellets in DMEM with 10% FBS （serum-containing group） and DMEM with 1% ITS, 10-7 M Dexamethasone, 50μg/mL Vitamin C （serum-free group）. Chondrocytes proliferation, matrix secretion and gene expression were tested by MTT assay, histological staining, immunohistochemist and semi-quantitative gene expression analysis. Results Serum-free culture medium could not promote chondrocytes proliferation and phenotype of chondrocytes could be maintained for only 3 days in monolayer culture. In 3-D culture, the histological, immunohistochemical results and expression of cartilage-specific genes were comparable between serum-containing group and serum-free group. Conclusion Serum-free culture medium could promote cartilage formation and maintain phenotype of chondrocytes in 3-D culture. Serum-free culture medium can be used for differential proteomics analysis and protein assay analysis.

关 键 词：无血清培养基 胰岛素铁硒传递蛋白 组织工程 软骨表型 

分 类 号：Q813.11[生物学—生物工程]