基材弹性模量对兔纤维环源干细胞和骨髓间充质干细胞分化调控的比较研究

发布时间：2018-04-30 15:21

本文选题：纤维环源干细胞 + 骨髓间充质干细胞　；参考：《苏州大学》2015年硕士论文

【摘要】：目的:分离培养兔纤维环源干细胞(Annulus Fibrosus-derived Stem Cells,AFSCs)和骨髓间充质干细胞(Bone marrow Mesenchymal Stem Cells,BMSCs),制备出具备不同弹性模量的聚氨酯静电纺丝纤维支架;探讨基材弹性模量对两种干细胞分化的调控,并比较两种干细胞在不同弹性模量聚氨酯静电纺丝纤维支架上向接近纤维环径向各区域细胞分化的能力。方法:获取同一只6月龄新西兰大耳白兔的纤维环和骨髓,分别分离培养纤维环源干细胞和骨髓间充质干细胞;通过静电纺丝法制备出不同弹性模量的聚氨酯静电纺丝纤维支架。分别将AFSCs和TGF-β3处理的BMSCs(t BMSCs)种植到支架材料上,通过MTS法测定两种干细胞在支架材料上的增殖能力,通过RT-q PCR和CTFM比较两种干细胞在支架材料上向接近纤维环径向各区域细胞分化的能力。结果:分离培养出兔AFSCs和BMSCs,两种干细胞均具有干细胞表面标记物OCT-4,Nucelostemin(NS)和SSEA-4表达;具有成脂,成骨及成软骨多向诱导分化能力。制备出具有不同弹性模量的聚氨酯静电纺丝纤维支架,具有较高的孔径率。MTS检测显示AFSC和t BMSCs在各组支架上均能增殖;在AFSCs和t BMSCs中,collagen-I的表达量随着基材弹性模量的增大而增高,而collagen-II和aggrecan的表达趋势相反;对于同一弹性模量的聚氨酯静电纺丝纤维支架而言,t BMSCs的collagen-I,collagen-II和aggrecan基因的表达量均高于AFSCs。此外,AFSCs和t BMSCs细胞牵引力都随着基材弹性模量的增加而降低,这与实际纤维环组织由内区到外区细胞牵引力的变化趋势相似。结论:兔AFSCs和t BMSCs细胞基因表达和细胞力学均受支架弹性模量的调控,具有相似的向接近纤维环径向各区域细胞分化的能力。
[Abstract]:Objective: to isolate and culture Annulus Fibrosus-derived Stem cells from rabbit fibroblast cells and bone marrow mesenchymal stem cells (BMSCs) to prepare polyurethane electrostatic spinning fiber scaffolds with different elastic moduli, and to investigate the regulation of elastic modulus of substrates on differentiation of two kinds of stem cells. The ability of the two stem cells to differentiate into the radial region of the fiber ring on the polyurethane fiber scaffold with different elastic modulus was compared. Methods: the fibrous ring and bone marrow of the same 6-month-old New Zealand white rabbit were isolated and cultured, and the bone marrow mesenchymal stem cells (BMSCs) and fibroring derived stem cells (FRSCs) were isolated and cultured respectively. Polyurethane fiber scaffolds with different elastic modulus were prepared by electrostatic spinning. The BMSCs(t BMSCs treated with AFSCs and TGF- 尾 3 were implanted into the scaffold material, and the proliferation ability of the two stem cells on the scaffold material was measured by MTS method. RT-q PCR and CTFM were used to compare the ability of the two stem cells to differentiate into the radial region of the fibrous ring on the scaffold material. Results: rabbit AFSCs and BMSCs were isolated and cultured. Both stem cells had the expression of OCT-4 Nucelosteminus NSN and SSEA-4, and had the ability of lipogenesis, osteogenesis and cartilage differentiation. Polyurethane electrospun fiber scaffolds with different elastic modulus were prepared. The results showed that AFSC and t BMSCs could proliferate on each group of scaffolds. The expression of collagen-I in AFSCs and t BMSCs increased with the increase of elastic modulus of the substrate, but the expression trend of collagen-II and aggrecan was opposite, and the expression of collagen-Igna collagen-II and aggrecan genes in t BMSCs was higher than that of AFSCs in the same elastic modulus polyurethane fiber scaffold. In addition, the tractive force of AFSCs and t BMSCs cells decreased with the increase of the elastic modulus of the substrate, which was similar to the change trend of the actual fibrous ring tissue from the inner region to the outer region. Conclusion: the gene expression and cellular mechanics of rabbit AFSCs and t BMSCs cells are regulated by the elastic modulus of scaffold and have the similar ability to differentiate into the radial region of the fibrous ring.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R681.5

【相似文献】