脂肪干细胞与纤维环细胞共培养后类纤维环分化效应的研究.pdfVIP

中国细胞生物学学报 Chinese Journal of Cell Biology 2013, 35(6): 797–803 http ://www.cj DOI: 10.11844/CJCB.2013.06.0043 脂肪干细胞与纤维环细胞共培养后的 类纤维环分化效应研究 胡旭琪 张小磊 郑旭浩 吴 畏 江立波 倪 晓 余 洋 周一飞 徐华梓* (温州医学院附属第二医院, 温州325000) 摘要 干细胞移植是治疗椎间盘退变的新方向, 而对纤维环的修复是治疗椎间盘退变的前 提。该研究利用Transwell共培养系统在常氧(20%)及低氧(2%)条件下探讨共培养诱导后SD 大鼠脂 肪干细胞向类纤维环细胞的分化能力。1,9-Dimethylmethylene blue(DMMB)检测表明, 低氧组脂肪 干细胞诱导7, 14, 2 1 d后及常氧组脂肪干细胞诱导14 d及2 1 d后与对照组脂肪干细胞相比分泌氨基 聚糖明显增加(P0.05), 且共培养14 d及2 1 d后低氧组脂肪干细胞较常氧组脂肪干细胞分泌氨基聚 糖含量明显增多(P0.05) 。qRT-PCR检测结果表明, 与对照组相比, 共培养14 d及2 1 d后常氧组及低 氧组ADSCs COL I、COL II及Aggrecan mRNA表达均明显增强(P0.05), 且低氧组ADSCs表达量高 于常氧组(P0.05) 。COL I 、COL II免疫组化及阿利辛染色也证实了上述结果。由此说明共培养可 以诱导脂肪干细胞向类纤维环细胞分化, 且低氧(2%)有利于这种分化的发生。 关键词 脂肪干细胞; 纤维环细胞; 共培养; 椎间盘退变 In vitro Differentiation of Adipose-derived Mesenchymal Stem Cells toward Annulus Fibrosus-like Cells by Co-culturing with Anulus Fibrosus Cells Hu Xuqi, Zhang Xiaolei, Zheng Xuhao, Wu Wei, Jiang Libo, Ni Xiao, Yu Yang, Zhou Yifei, Xu Huazi* (The Second Affiliated Hospital of Wenzhou Medical College, Wenzhou 325000, China) Abstract In this research, we studied the differentiation ability of adipose-derived mesenchymal stem cells (ADSCs) toward annulus fibrosus-like cells by co-culturing with annulus fibrosus cells (AFCs) under nor- moxia (20% O ) or hypoxia (2% O ) respectively. 1,9-Dimethylmethylene blue (DMMB) assay found glycosamin- 2 2 oglycan synthesis of hypoxia group which co-cultured for 7, 14, 21 d and normoxia group which co-coltured for 14, 21 d was significantly upregulated (P0.05). The ADSCs in hypoxia group producted more glycosaminogly- can than those in normoxia group (P0.05). The expression of collagen type I (COL I), collagen typ