体外共培养软骨细胞与脂肪基质细胞用于软骨构建实验研究.docVIP

体外共培养软骨细胞与脂肪基质细胞用于软骨构建的实验研究 　　[摘要] 目的 探讨体外共培养软骨细胞与脂肪基质细胞（ADSCs）用于软骨构建的可行性。 方法 分别收集并培养人ADSCs与猪耳软骨细胞，设置实验组、阳性对照组、阴性对照组，分别接种ADSCs和软骨细胞（以7∶3比例混合）、单纯软骨细胞、单纯ADSCs，观察并对比三组的形态学变化、湿重、蛋白多糖含量、组织学特征及Ⅱ型胶原的表达情况。 结果 经过8周的体外培养，实验组组织形状规则，表现出软骨组织的结构特征，且有一定的弹性；对于平均湿重及蛋白多糖定量检测发现，实验组的平均湿重可达阳性对照组的73.1%、81.9%，与阴性对照组比较差异有统计学意义（P0.01）；HE染色显示实验组标本出现连续的软骨样组织，产生成熟的软骨陷窝及纤维性组织，新生软骨厚度较为明显，Ⅱ型胶原免疫组化染色发现，实验组软骨陷窝附近多处呈棕黄色，呈阳性反应。结论 软骨细胞及ADSCs体外共培养可用于软骨组织的构建，还需进一步研究确定ADSCs转化为成熟软骨细胞的直接证据。 　　[关键词] 软骨细胞；脂肪基质细胞；共培养；软骨构建 　　[中图分类号] R321[文献标识码] A[文章编号] 1674-4721（2014）06（c）-0012-04 　　 　　Experimental study of in vitro co-culture of chondrocytes and adipose-derived stromal cells for cartilage construction 　　JIA Li CUI Jun 　　Department of Stomatology,Peking University Shenzhen Hospital,Shenzhen 518000,China 　　[Abstract] Objective To investigate the feasibility of in vitro co-culture of chondrocytes and adipose-derived stromal cells (ADSCs) for cartilage construction. Methods ADSCs and porcine auricular chomdrocytes were collected and cultured in vitro,and then three groups were set as the experimental group,the positive control group and the negative control group,which were inoculated ADSCs and chondrocytes(7∶3 mixing ratio),simple chondrocytes,simply ADSCs respectively.And the contrast morphological changes,the wet weight,the proteoglycan content changes and type Ⅱ collagen in the expression of histological feature of the three groups was observed and analyzed respectively. Results After eight weeks in vitro culture,the tissue of experimental group had a regular shape,which looked like the structure of cartilage tissue and was certain flexibility.For detection ofthe average wet weight and proteoglycan quantitative,the average wet weight and proteoglycan could reach 73.1%,81.9% of that in the positive experimental group respectively,which were significantly higher than that in the negative control group(P0.01).HE staining showed that the experimental group occurred consecutive cartilage-like tissue,matur