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《不同方法誘導(dǎo)去分化脂肪細(xì)胞心肌樣細(xì)胞分化的實(shí)驗(yàn)分析》由會(huì)員上傳分享,免費(fèi)在線閱讀,更多相關(guān)內(nèi)容在學(xué)術(shù)論文-天天文庫。
1、-------NRHCL組(P<0.05)。結(jié)論:①天花板貼壁培養(yǎng)法可成功的使成熟脂肪細(xì)胞去分化為DFAT細(xì)胞,DFAT細(xì)胞經(jīng)乳鼠心臟組織裂解液、維生素C體外誘導(dǎo)分化后可再分化為心肌樣細(xì)胞,是心肌細(xì)胞再生較為理想的種子細(xì)胞來源。②維生素C可以提高乳鼠心臟組織裂解液體外誘導(dǎo)DFAT細(xì)胞向心肌分化的效率。③與對照組相比,DFAT細(xì)胞經(jīng)維生素C、乳鼠心臟組織裂解液誘導(dǎo)后心肌特異性標(biāo)志物的mRNA和蛋白質(zhì)表達(dá)水平均升高。本實(shí)驗(yàn)尚不能證實(shí)二者誘導(dǎo)能力有差異,二者聯(lián)合應(yīng)用可使誘導(dǎo)能力可增強(qiáng)(P<0.05)。但均未觀察到有自發(fā)跳動(dòng)的細(xì)胞。故本實(shí)驗(yàn)只
2、誘導(dǎo)出心肌樣心肌前體細(xì)胞,離誘導(dǎo)出功能心肌細(xì)胞還有較大的距離?!娟P(guān)鍵詞】去分化脂肪細(xì)胞維生素C心臟組織裂解液誘導(dǎo)分化III-----------EffectsofDifferentInductionConditionsonDifferentiationofDedifferentiatedFatCellsintoCardiomyocyte-likeCellsMajor:MedicinecardiovascularPostgraduate:FuhaiLiSupervisor:ChenyunZhang[Abstract]Objective:
3、Tostudytheabilityofdifferentmethodsininducingdifferentiatedfatcells(dedifferentiatedfatcells,DFAT)todifferentiatetowardscardiomyocytesinvitroandthedifferencesofmyocardialtumor-specificmarkersexpression,andtooptimizeconditionsininducingdifferentiationofDFATcellsintocardi
4、omyocytes,inordertoprovideexperimentalfoundationtoimproveseedcellsforacutemyocardialinfarctionincelltherapy.Method:Extractmaturefatcellsfromadiposetissueofadultrats(matureadipocytes,MA)anddedifferentiatethemintoDFATcellsbyceilingadherentculturemethod,thenculturethethird
5、-generationDFATcellswithmediumcontainingthevitaminC(GroupI),orneonatalratheartcelllysate(GroupII),orbothvitaminCandneonatalratheartcelllysate(GroupIII),orordinarycompletelymedium(GroupIV,controlgroup)forthreeweeksinductiontraining,thenobservethechangesofcellmorphology.C
6、arryoutcTnTimmunofluorescencetoidentifyDFATcellofthreeweeksinduction,andreal-timePCRandwesternblottingtodetecttheexpressionofmyocardialspecificmarkerscTnT,GATA-4andNKx2.5ofeachgroupinmRNAandproteinlevels.Results:Aftertheceilingadherentculturing,MAdifferentiatedintoDFATc
7、ellswithfibroblastsmorphologyinvitro.AftervitaminCinduction,neonatalratheartcelllysateinductionorthejointinduction,DFATcellswasobservedtostretchthecellbodyintospindleshape,enlargetheirsizeandprotrudebranchingprocesses,andconnectwithothercellstoformcellclusters,myotube-l
8、ikestructurewasobservedandcellmorphologywassimilartomyocardialcells,butnospontaneousbeatingcellswereobserved.I