摘要
[目的]通过非接触式共培养探索兔髓核间充质干细胞(NPMSCs)与兔髓核细胞(NPCs)的间接生物学效应.[方法]将第3代兔来源NPMSCs与NPCs进行非接触共培养,分设三组:NPCs/NPCs自身共培养对照组,NPMSCs/NPMSCs自身共培养对照组,NPMSCs/NPCs共培养组.分别在共培养3、5、7d后,比较两种细胞增殖情况,ELISA检测上清液中转化生长因子β1(TGF-β1)、胰岛素样生长因子(IGF)的含量;采用RT-PCR检测共培养7d后NPMSCs与NPCs的Col Ⅱc1、AGG基因表达变化;免疫荧光染色检测NPMSCs中Col Ⅱ的蛋白表达情况.[结果]在共培养第5d和第7d,共培养组NPCs的细胞数量均高于自身对照组(P<0.05),在第7d,共培养组NPMSCs的细胞数量均高于自身对照组;在共培养3、5、7d后,共培养组上清液TGF-β1、IGF的含量均高于自身对照组(P<0.05);在培养7d后,共培养组NPMSCs中Col Ⅱ免疫荧光强度高于自身对照组,NPCs与NPMSCs的Col Ⅱa1、AGG基因的表达较自身对照组显著升高(P<0.05).[结论]NPMSCs与NPCs非接触共培养可以促进细胞因子分泌、细胞增殖、基质合成与分泌.
[Objective] To explore the indirect biological effects of non-contact coculture between rabbit-derived nucleus pulposus mesenchymal stem cells(NPMSCs) and rabbit nucleus pulposus cells(NPCs).[Method] The P3 generation rabbit-derived NPMSCs were co-cultured with NPCs by non-contacted ways, which were divided into three groups: NPCs/NPCs, NPMSCs/NPMSCs and NPMSCs/NPCs co-culture group. After 3, 5, and 7 days of co-culture, proliferation capacity of the two type of cells was tested. The content of transforming growth factor β1(TGF-β1) and insulin-like growth factor(IGF) in the supernatant was detected by ELISA. In addition, the gene expressions of Col IIα1 and AGG in NPMSCs and NPCs 7 days after co-culture were detected by using RT-PCR, while the protein expression of Col II in NPMSCs was assessed by immunofluorescence staining.[Results] At 5 and 7 days after coculture, the number of NPCs in the co-culture group was significantly higher than that in other control groups(P<0.05). Similarly, at 7 days of coculture, the number of NPMSCs in the coculture group was higher than that in other control groups. In addition, the content of TGF-β1 and IGF in the supernatant of the co-culture group was significantly higher than those in other two control groups at 3, 5 and 7 days(P<0.05). At 7 days, the intensity immunofluorescence of Col II was higher than that of control group, whereas the gene expression of Col IIα1 and AGG in NPCs and NPMSCs was significantly higher than that in control group(P<0.05).[Conclusion] Non-contact coculture of NPMSCs with NPCs do promote cytokine secretion, cell proliferation, matrix synthesis and secretion.
作者
陈晓峰
郭伟俊
韩桩汛
褚学远
李浩
劳泽辉
欧志聪
蔡东岭
CHEN Xiao-feng;GUO Wei-jun;HAN Zhuang-xun;CHU Xue-yuan;LI Hao;LAO Ze-hui;OU Zhi-cong;CAI Dong-ling(Department of Orthopaedics,Panyu Hospital of Chinese Medicine,Guangzhou 511400,China)
出处
《中国矫形外科杂志》
CAS
CSCD
北大核心
2019年第11期1018-1024,共7页
Orthopedic Journal of China
基金
广东省中医药局科研项目(编号:20191262)
