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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。
Stem cell transplantation is a new direction for the treatment of disc degeneration, and the repair of annulus fibrosus is a prerequisite for the treatment of disc degeneration. In this study, Transwell co-culture system was used to investigate the differentiation of ADSCs into fibroblasts after induction of co-culture under normoxia (20%) and hypoxia (2%) conditions. The results of 1, 9-Dimethylmethylene blue (DMMB) showed that the adipose-derived stem cells of hypoxia group were significantly less than those of adipose-derived stem cells on the 7th, 14th, and 21st days after inducing, (P <0.05). Compared with the normoxia group, adipose-derived stem cells in the hypoxia group had significantly higher levels of aminoglycan (P <0.05) on the 14th and 21st day after co-culture. The results of qRT-PCR showed that the expression of COL I, COL II and Aggrecan mRNA of ADSCs in normoxia group and hypoxia group were significantly increased (P <0.05) at 14 d and 21 d after co-culture compared with the control group, and hypoxia Group ADSCs expression was higher than normoxic group (P <0.05). COL I, COL II immunohistochemistry and Aliskiren staining also confirmed the above results. Thus co-culture can induce adipose-derived stem cells to differentiate into fibrocystic ring cells, and hypoxia (2%) is conducive to the occurrence of this differentiation.