發(fā)布時間：2018-04-30 15:21

  本文選題：纖維環(huán)源干細胞 + 骨髓間充質(zhì)干細胞��； 參考：《蘇州大學(xué)》2015年碩士論文

【摘要】：目的:分離培養(yǎng)兔纖維環(huán)源干細胞(Annulus Fibrosus-derived Stem Cells,AFSCs)和骨髓間充質(zhì)干細胞(Bone marrow Mesenchymal Stem Cells,BMSCs),制備出具備不同彈性模量的聚氨酯靜電紡絲纖維支架;探討基材彈性模量對兩種干細胞分化的調(diào)控,并比較兩種干細胞在不同彈性模量聚氨酯靜電紡絲纖維支架上向接近纖維環(huán)徑向各區(qū)域細胞分化的能力。方法:獲取同一只6月齡新西蘭大耳白兔的纖維環(huán)和骨髓,分別分離培養(yǎng)纖維環(huán)源干細胞和骨髓間充質(zhì)干細胞;通過靜電紡絲法制備出不同彈性模量的聚氨酯靜電紡絲纖維支架。分別將AFSCs和TGF-β3處理的BMSCs(t BMSCs)種植到支架材料上,通過MTS法測定兩種干細胞在支架材料上的增殖能力,通過RT-q PCR和CTFM比較兩種干細胞在支架材料上向接近纖維環(huán)徑向各區(qū)域細胞分化的能力。結(jié)果:分離培養(yǎng)出兔AFSCs和BMSCs,兩種干細胞均具有干細胞表面標(biāo)記物OCT-4,Nucelostemin(NS)和SSEA-4表達;具有成脂,成骨及成軟骨多向誘導(dǎo)分化能力。制備出具有不同彈性模量的聚氨酯靜電紡絲纖維支架,具有較高的孔徑率。MTS檢測顯示AFSC和t BMSCs在各組支架上均能增殖;在AFSCs和t BMSCs中,collagen-I的表達量隨著基材彈性模量的增大而增高,而collagen-II和aggrecan的表達趨勢相反;對于同一彈性模量的聚氨酯靜電紡絲纖維支架而言,t BMSCs的collagen-I,collagen-II和aggrecan基因的表達量均高于AFSCs。此外,AFSCs和t BMSCs細胞牽引力都隨著基材彈性模量的增加而降低,這與實際纖維環(huán)組織由內(nèi)區(qū)到外區(qū)細胞牽引力的變化趨勢相似。結(jié)論:兔AFSCs和t BMSCs細胞基因表達和細胞力學(xué)均受支架彈性模量的調(diào)控,具有相似的向接近纖維環(huán)徑向各區(qū)域細胞分化的能力。
[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.
【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R681.5

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