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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)-研究》由會(huì)員上傳分享，免費(fèi)在線閱讀，更多相關(guān)內(nèi)容在應(yīng)用文檔-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(duì)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
2、動(dòng)的細(xì)胞。故本實(shí)驗(yàn)只誘導(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。【關(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萬方數(shù)據(jù)EffectsofDifferentInductionConditionsonDifferentiationofDedifferentiatedFatCellsintoCardiomyocyte-likeCellsMajor:MedicinecardiovascularPostgraduate:FuhaiLiSupervisor:ChenyunZhang
3、[Abstract]Objective：Tostudytheabilityofdifferentmethodsininducingdifferentiatedfatcells(dedifferentiatedfatcells,DFAT)todifferentiatetowardscardiomyocytesinvitroandthedifferencesofmyocardialtumor-specificmarkersexpression,andtooptimizeconditionsininduci
4、ngdifferentiationofDFATcellsintocardiomyocytes,inordertoprovideexperimentalfoundationtoimproveseedcellsforacutemyocardialinfarctionincelltherapy.Method：Extractmaturefatcellsfromadiposetissueofadultrats(matureadipocytes,MA)anddedifferentiatethemintoDFATc
5、ellsbyceilingadherentculturemethod,thenculturethethird-generationDFATcellswithmediumcontainingthevitaminC(GroupI),orneonatalratheartcelllysate(GroupII),orbothvitaminCandneonatalratheartcelllysate(GroupIII),orordinarycompletelymedium(GroupIV,controlgroup
6、)forthreeweeksinductiontraining,thenobservethechangesofcellmorphology.CarryoutcTnTimmunofluorescencetoidentifyDFATcellofthreeweeksinduction,andreal-timePCRandwesternblottingtodetecttheexpressionofmyocardialspecificmarkerscTnT,GATA-4andNKx2.5ofeachgroupi
7、nmRNAandproteinlevels.Results：Aftertheceilingadherentculturing,MAdifferentiatedintoDFATcellswithfibroblastsmorphologyinvitro.AftervitaminCinduction,neonatalratheartcelllysateinductionorthejointinduction,DFATcellswasobservedtostretchthecellbodyintospindl
8、eshape,enlargetheirsizeandprotrudebranchingprocesses,andconnectwithothercellstoformcellclusters,myotube-likestructurewasobservedandcellmorphologywassimilartomyocardialcells,butnospontaneousbeatingcellswereobserved.Immunofluoresce
