资源描述：
《不同方法诱导去分化脂肪细胞心肌样细胞分化的实验-研究》由会员上传分享，免费在线阅读，更多相关内容在应用文档-天天文库。
1、NRHCL组(P<0.05)。结论：①天花板贴壁培养法可成功的使成熟脂肪细胞去分化为DFAT细胞，DFAT细胞经乳鼠心脏组织裂解液、维生素C体外诱导分化后可再分化为心肌样细胞，是心肌细胞再生较为理想的种子细胞来源。②维生素C可以提高乳鼠心脏组织裂解液体外诱导DFAT细胞向心肌分化的效率。③与对照组相比，DFAT细胞经维生素C、乳鼠心脏组织裂解液诱导后心肌特异性标志物的mRNA和蛋白质表达水平均升高。本实验尚不能证实二者诱导能力有差异，二者联合应用可使诱导能力可增强(P<0.05)。但均未观察到有自发跳
2、动的细胞。故本实验只诱导出心肌样心肌前体细胞，离诱导出功能心肌细胞还有较大的距离。【关键词】去分化脂肪细胞维生素C心脏组织裂解液诱导分化III万方数据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
