自体软骨细胞复合Ⅰ型胶原支架体外动态培养的实验研究

发布时间：2018-03-19 08:25

  本文选题：软骨细胞　切入点：Ⅰ型胶原支架材料　出处：《中国矫形外科杂志》2015年18期 　论文类型：期刊论文

【摘要】：[目的]比较体外静态与动态培养两种方式对培养人关节软骨细胞复合Ⅰ型胶原支架材料的效果,从而探索体外构建移植物用于治疗关节软骨缺损的适宜条件及方式。[方法]分离培养人关节软骨细胞,P2代软骨细胞接种于Ⅰ型胶原支架,随机分为3组,A组:静态培养;B组:动态培养;C组:单层培养。采用倒置相差显微镜、荧光显微镜、SEM、HE染色观察细胞在支架上的分布、黏附、生长及形态特点,实时荧光定量PCR检测软骨细胞表型特异基因Ⅱ型胶原的表达情况。[结果]体外单层培养的软骨细胞(P2)以多角形为主,Ⅱ型胶原蛋白表达阳性,提示P2代细胞表型维持良好;与A组比较,B组细胞数量较多且分布均匀,细胞形态较均一,以多角形为主;A、B组样本大体结构及内部孔隙结构均保持完整,孔隙内均可见细胞黏附,B组孔隙内细胞数量及细胞外基质优于A组;A、B组的Ⅱ型胶原基因mRNA表达水平较单层培养组明显增加。[结论]软骨细胞复合Ⅰ型胶原支架体外动态培养较静态培养可明显促进细胞增殖及细胞外基质分泌。因此,动态培养有望成为体外构建自体软骨细胞移植术所需移植物的一种有效方法。
[Abstract]:[objective] to compare the effects of static and dynamic culture in vitro on cultured human articular chondrocytes combined with type I collagen scaffolds. Therefore, to explore the suitable conditions and methods of constructing grafts for the treatment of articular cartilage defects in vitro. [methods] chondrocytes of P2 generation were isolated from cultured human articular chondrocytes and inoculated on type I collagen scaffolds. Three groups were randomly divided into three groups: static culture group B: dynamic culture group C: monolayer culture. The distribution, adhesion, growth and morphological characteristics of the cells on the scaffold were observed by inverted phase contrast microscope and fluorescence microscope SEMHE staining. The expression of phenotypic specific gene type 鈪，

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