bdnf基因修饰施万细胞结合组织工程技术治疗周围神经损伤的分析word格式论文.docxVIP

织贴壁培养方法快,进一步纯化后做S-100 细胞免疫组织化学鉴定为SCs，纯度达到95%以上。（2）SCs与壳聚糖支架的组织相容性：正常对照组和实验组各时点凋亡细胞的百分率经差异无显著性意义(P0.05)。（3）大体观察：饲养过程中无大鼠死亡，大鼠患肢屈曲挛缩情况及拖曳步态在6周以后有所改善。12周后大鼠处死，未见导管残留及桥接段上下神经萎缩，周围有血管增生、粘连现象。（4）电生理检测：四组NCV相互比较，A组与B组、A组与C组均有显著性差异(P0.05)，A组与D组无显著性差异(P0.05)。组织学观察组织工程神经体内长期、稳定表达目的基因-BDNF和报告基因-GFP、FLAG。结论:（1）先用酶消化,后组织块贴壁培养SCs和差速贴壁综合运用的纯化方法所“生产”的SCs数量较大,纯度较高,且细胞的理化性质及生物学特性受外界环境影响较小。同时又节约了神经组织的材料来源,避免了反复多次取材的繁琐操作过程,节省人力和物力,并且保证了实验所需的SCs尽可能来源于同一只或同一批动物。（2）使用冷冻干燥技术研制的壳聚糖导管表面光滑，透明状，质地均匀。厚度0.15mm，与盖玻片的厚度相仿。实验表明具有良好生物相容性，SCs可以在其表面生长增殖。可以作为神经桥接管。（3）用壳聚糖制作三维支架，以BDNF 基因转染的SCs为种子细胞，构建成的组织工程化人工神经，修复神经缺损，其修复效果接近于自体神经移植，相对于单纯应用壳聚糖神经导管修复神经缺损可进一步提高神经修复的效果。具有广阔的应用前景，值的进一步研究。关键词:施万细胞BDNF神经生长因子周围神经再生壳聚糖EffectsofBDNF-modifiedSchwanncellsandtissueengineeringinthetreatmentofperipheralnerveinjuryHumanAnatomy and Embryology Zhang FengjiuDirected by Zhao ZhiyingAbstractObjectine:(1)Researchingexternalhigh-purity“Wistar”culturalmethodof sucklingratcells.(2)InsertingBDNFschwanncellintopresettingChitosantubes,constructingtissueengineeringartificialnervousbridgeconnectrat10mmSCssciaticnervedamnify,researchingtherepairingeffectofartificialnervousbridgeandthese interactionbetweennerveregenerationfactor,providingexperimentevidencefor tissue engineeringfurtherstudies and clinical application.Method:(1)Take3-5dayssucklingrattwosidessciaticnerveandbrachial plexus,afterdigestingbyenzyme,explantsplantinginsolatingcultivateSCs.Aftera week’scultivation,fullofbottom,adding0.25%trypsasegeneraldigestedtoeach bottlestogeneration,usingS-100immunohistochemistrydyeingandcalculatingSCs purity.(2)Getsuchschwanncellandmakethemintosuspension,densityis1×105/ml a24holecostar,dividedintonormalgroupandexperimentalgroup,Chitosantileson thebottomofholes,ultra-cleanplatformdrying.SCsvaccinatesintocostar, cultivatingisintheconditionof37 oCand5%CO2After24h,addingDAPI(final concentration 50μg/ml) intoincubation , After24h,washingbottleusingPBSat least3 times,toberidofDAPIwhichisnotcombinedwithcells.Addinginoculumsto continuecultivate.