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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胞去分化為DFAT細胞,DFAT細胞經(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胞,是心肌細胞再生較為理想的種子細胞來源。②維生素C可以提高乳鼠心臟組織裂解液體外誘導(dǎo)DFAT細胞向心肌分化的效率。③與對照組相比,DFAT細胞經(jīng)維生素C、乳鼠心臟組織裂解液誘導(dǎo)后心肌特異性標志物的mRNA和蛋白質(zhì)表達水平均升高。本實驗尚不能證實二者誘導(dǎo)能力有差異,二者聯(lián)合應(yīng)用可使誘導(dǎo)能力可增強(P<0.05)。但均未觀察到有自發(fā)跳
2、動的細胞。故本實驗只誘導(dǎo)出心肌樣心肌前體細胞,離誘導(dǎo)出功能心肌細胞還有較大的距離?!娟P(guān)鍵詞】去分化脂肪細胞維生素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