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  

In vitro Differentiation of Adipose-derived Mesenchymal Stem Cells toward Annulus Fibrosus-like Cells by Co-culturing with Anulus Fibrosus Cells

作　　者：胡旭琪[1] 张小磊[1] 郑旭浩[1] 吴畏[1] 江立波[1] 倪晓[1] 余洋[1] 周一飞[1] 徐华梓[1] 

机构地区：[1]温州医学院附属第二医院,温州325000

出　　处：《中国细胞生物学学报》2013年第6期797-803,共7页Chinese Journal of Cell Biology

基　　金：浙江省"重中之重"学科开放基金(批准号:2011GK001);温州市科学技术局资助项目(批准号:S20100048;Y20100357;Y20100211)资助的课题~~

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7,14,21 d后及常氧组脂肪干细胞诱导14 d及21 d后与对照组脂肪干细胞相比分泌氨基聚糖明显增加(P<0.05),且共培养14 d及21 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(P<0.05)。qRT-PCR检测结果表明,与对照组相比,共培养14 d及21 d后常氧组及低氧组ADSCs COL I、COL II及Aggrecan mRNA表达均明显增强(P<0.05),且低氧组ADSCs表达量高于常氧组(P<0.05)。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。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 normoxia （20% 02） or hypoxia （2% O2） respectively. 1,9-Dimethylmethylene blue （DMMB） assay found glycosaminoglycan 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 （P〈0.05）. The ADSCs in hypoxia group producted more glycosaminoglycan than those in normoxia group （P〈0.05）. The expression of collagen type I （COL I）, collagen type II （COL I1） and Aggrecan was estimated by quantitative reverse transcription-polymerase chain reaction. After 14 d and 21 d of co-culture, the expression of COL I, COL II and Aggrecan was significantly upregulated in hypoxia group and nomorxia group（P〈0.05）. And ADSCs in hypoxia group demonstrated more COL I, COL II and Aggrecan expression than those in nomorxia group （P〈0.05）. These results suggested that with a co-culture system, ADSCs can differentiate into annulus fibrosus-like cells. Further, the oxygen concentration significantly influenced the ADSCs differentiation.

关 键 词：脂肪干细胞 纤维环细胞 共培养 椎间盘退变 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]