学科分类
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11 个结果
  • 简介:Objective:Toexplorethecultureconditionsofhumanneuralstemcellsandtoinvestigatetheultrastructureofneurospheres.Methods:Thecellsfromtheembryonichumancorticesweremechanicallydissociated.N2mediumwasadaptedtocultureandexpandthecells.ThecellswereidentifiedbyimmunocytochemistryandEMwasappliedtoexaminetheultrastructureofneurospheres.Results:Theneuralstemcellsfromhumanembryonicbrainsweresuccessfullyculturedandformedtypicalneurospheresinsuspension,andmostofthecellsexpressedvimentin,whichwasamarkerforneuralprogenitorcells,andthecellscoulddifferentiateintoneurons,astrocytesandoligodendrocytes.Invitromyelinformationinneurosphereswereobservedatanearlystageofculture.Conclusions:Humanneuralstemcellscanbeculturedfromembryonicbrains,canformthetypicalneurospheresinsuspensioninvitroandhavetheabilityofmyelinating,andmaybepotentialsourcefortransplantationintreatingmyelindisorders.

  • 标签: 髓磷酯 神经半球 神经干细胞 细胞培养 细胞移植
  • 简介:客观:为了探索可行性构造遗传工程人,神经干细胞(hNSCs)由lentivirus调停了多表示基因以便为针的绳索损害(SCI)的进一步的研究提供接枝来源。方法:从人的流产胎的大脑外皮的人的神经干细胞被孤立并且有教养,然后,基因被lentivirus修改两个都表示绿荧光蛋白质(GFP)和老鼠neurotrophin-3(NT-3);转基因的表示被荧光显微镜,胎儿的老鼠的背面的根中心和槽污点的方法检测。结果:遗传工程hNSCs成功地被构造。所有在荧光显微镜下面表示了明亮绿的荧光的遗传工程hNSCs被观察。转基因的hNSCs的调节媒介能导致从背面的根中心(DRG)挥舞长出的神经突。遗传工程hNSCs表示了高级NT-3which能被使用槽污点检测。结论:遗传工程hNSCs调停了bylentivirus能被构造多成功地表示基因。

  • 标签: 遗传因素 实验研究 神经干细胞 多基因
  • 简介:客观:为了由adipose-derivedstromal房间(ASC)在vitro导致adipocyte区别,与绿荧光灯的蛋白质(GFP)从转基因的老鼠收获了并且估计经由ASC的附件构造脂肪质的纸巾打字的可能性我骨胶原scaffolds.Methods:从GFP转基因的老鼠的腹股沟的胖垫被酶为ASC(主要文化)的隔离消化。在到ASC的三个段落的扩大以后,房间被ASC在vitro在为二个星期,和adipocyte区别中等的anadipogenic孵化被词法观察和油红估计染色的O。然后,他们被纳入为12个小时andco有教养的骨胶原脚手架,为2months由皮下注射的培植列在后面到裸体老鼠的背面的皮肤。最新形成的纸巾被检测由他染色。结果:有教养的主要干细胞是像成纤维细胞、显示出的活跃增长。在在anadipocyte区别媒介被孵化以后,在逐渐地并且最后积累的细胞质的类脂化合物微滴发展成成熟adipocytes,它出现了在染色的油红O积极。0。5厘米的~3new组织块在裸体老鼠的背面的皮下面被发现,它被荧光灯的观察作为成熟脂肪质的纸巾证实并且他染色。结论:ASC能成功地区分脂肪质的纸巾进成熟adipocytes,它作为intracytoplasmic类脂化合物微滴展出象adipocyte一样形态学和快车。它是与ASC和类型设计的脂肪质的纸巾的一个有效模特儿我骨胶原脚手架。

  • 标签: 脂肪组织 干细胞 病理 治疗 临床
  • 简介:ObjectToinvestigatetheclinicalsignificanceofallogeneichematopoieticstemcelltransplantation(allo-HSCT)followingfludarabine(Flu)-basednomnyeloablativeconditioningregimen.Methods7patientswitholderageororgandysfunctionreceivedeitheroftwoFlu-basednonmyeloablativeconditioningprotocolsfollowedbyinfusionofgranulocytecolony-stimulatingfactor(G-CSF)mobilizedallogeneicperipheralblooclstemcells(PBSC).Cy-elosporincombiningmethotrexatewasusedasgraftvshostdisease(GVHD)prophylaxis.ResultsMinimalalloHSCTassociatedtoxicitywasfoundapartfrommucositis.Theallogeneicdonorengraftmentswereverifiedinallthepatients.Sixofseveneasessurvivedmorethan5months.AcuteGVHDoccurredinthreeofsevenpatientsincludingacaseofgrade1IGVHD.ConclusionTherapidengrafunentofallo-PBSCandgraftvsleukemia(GVL)effectscanbeobtainedbyFlu-basednomnyeloablativeconditioningregimen.Thismanagementissuitableforthepatientswhoaretoooldorhaveorgandysfunction.

  • 标签: GVHD PBSC HSCT GVL CSF
  • 简介:探索骨头的效果的目的有带hepatocyte生长因素的adenoviral向量的导出髓的间充质的干细胞(BMSC)transfected(HGF,Ad-HGF)在灼伤创伤愈合上。从男Wistar老鼠的方法BMSC用由密度坡度centrifugation中等、与包含20%胎儿的牛的浆液(FBS)的DMEM有教养的Percoll分开被分开并且净化。当时,BMSC是有在感染(MOI)的100复合的最佳的基因transduction效率的Ad-HGF的transfected。transfection和在暂停的HGF的表示的效率被流动cytometry检测,酶分别地连接了immunosorbent试金(ELISA)。32只雌老鼠受到90慥?灳瑩吗?

  • 标签: 肝细胞生长因子 骨髓间质干细胞 创面愈合 基因转染 烧伤 WISTAR大鼠
  • 简介:Objective:Toinvestigatethedifferentiativecapabilityofadulthumanbonemarrowmesenchymalstemcells(BMSCs)intoSchwann-likecells.Methods:BonemarrowswereaspiratedfromhealthydonorsandmononuclearcellswereseparatedbyPercolllymphocytesseparationliquid(1.073g/ml)withcentrifugation,cellswereculturedinDMEM/F12(1:1)mediumcontaining10%fetalbovineserum(FBS),20ng/mlepidermalgrowthfactor(EGF)and20ng/mlbasicfibroblastgrowthfactor(bFGF).Cellsofpassage1wereidentifiedwithimmunocytochemistry.Conclusions:BonemarrowcontainsthestemcellswiththeabilityofdifferentiatingintoSchwann-likecells,whichmayrepresentanalternativestemcellsourcesforneuraltransplantation.

  • 标签: 成年 骨髓间充质干细胞 施沃恩样细胞 神经移植
  • 简介:Objective:Tofurtherinvestigatetheosteogenicpotentialofrabbitmarrowstromalstemcellsculturedinvitro.Methods:Rabbitmarrowstromalstemcellswereisolatedbydensitygradientcentrifugationmethodandamplifiedintheflasks,usingtheosteogenicinducingconditions(OGC)astheculturemedia.Theosteogenicpotentialofmarrowstromalstemcellswereinvestigatedbymeansofbone-seekingfluorescenc(tetracycline)labeling,AlizarinredS(ARS)staining,Alcianblue-Siriusred(AS)staining,andscanningelectronmicroscope.Results:Afterbeingpassaged,themarrowstromalstemcellsincreasedinnumber,becameconfluentandformedmulti-layerstructure.Thestromalstemcellsexcretedinnumerabletinygranules,heapinguponthecellbodyandmerginggraduallyintofoggysubstances.Thesefoggysubstanceskeptonenlargingandformedround,oval,orflake-likenodules.Thesenodulesrevealedbrightgoldenyellowfluorescenceunderfluorescencemicroscopewhenlabeledwithtetracycline.HistochemicalstudywithspecificnewbonestainingwithARSrevealedpositivecalciumreaction,bothdenotingthattheywerenewlyformedbonetissues.AftertheywerestainedwithAS,collagenandacidmucopolysaccharidewereshown.Underscanningelectronmicroscope,threetypesofcellswithdifferentconfigurationswerefound.Theywereglobularcells,spindle-shapedcellsandpolygonalorpolygonalcells.Granuleswereexcretedfromthecellsandheapeduponthecellbody.Needle-shapedandirregularlyrectangularcrystalsalsoappearedandagglomeratedwiththegranulestoformnodulesandtrabecula-likeorflake-likestructures.Conclusions:Sequenceofeventsofboneformationbyrabbitmarrowstromalstemcellsculturedinvitroisfullydepictedandconfirmed,whichprovidesthefoundationforfurtherinvestigatingthemechanismsofosteoblastdifferentiationfrommarrowstromalstemcellsandthepossibleapplicationinorthopaedics.

  • 标签: 家兔 骨髓基质干细胞 体外培养 成骨能力 组织化学 扫描电镜
  • 简介:Objective:Toevaluatetheosteogenicpotentialofbonemorphogeneticprotein(BMP)-2genetransfectedgoatbonemarrow-derivedmesenchymalstemcells(MSCs).Methods:Goatbonemarrow-derivedMSCsweretransfectedbyAdv-humanbonemorphogeneticprotein(hBMP)-2gene(Group1),Adv-betagaltransfectedMSCs(Group2)anduninfectedMSCs(Group3).Westernblotanalysis,alkalinephosphatasestaining,VonKossastainingandtransmissionelectronmicroscopywereadoptedtodeterminethephenotypeofMSCs.Thenthecellswereinjectedintothighmusclesofthenudemice.Radiographicalandhistologicalevaluationswereperformedatdifferentintervals.Results:OnlyAdv-hBMP-2transfectedMSCsproducedhBMP-2.Thesecellswerepositiveforalkalinephosphatasestainingatthe12thdayandwerepositiveforVonKossastainingatthe16thdayaftergenetransfer.Electronmicroscopicobservationshowedthatthereweremoreroughendoplasmicreticulum,mitochondriaandlysosomesinAdv-hBMP-2transfectedMSCscomparedtoMSCsofothertwogroups.Atthe3rdand6thweeksaftercellinjection,ectopicboneswereobservedinmusclesofnudemiceofGroup1.Onlyfibroustissueoralittlebonewasfoundinothertwogroups.Conclusions:BMP-2genetransfectedMSCscandifferentiateintoosteoblastsinvitroandinduceboneformationinvivo.

  • 标签: 异位骨形成 骨成形蛋白-2 基因转染 骨髓间充质干细胞 小鼠 动物实验