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时间:2020-06-03
《电刺激促骨髓间充质干细胞向心肌样细胞分化.pdf》由会员上传分享,免费在线阅读,更多相关内容在行业资料-天天文库。
1、第30卷第3期生物医学工程学杂志VoI.3ONo.32013年6月JournalofBiomedicalEngineeringJune20l3电刺激促骨髓间充质干细胞向心肌样细胞分化*严中琴杨刚崔燎。何学令况薇吴文超刘小菁李良△1(四川大学华西基础医学与法医学院,生物医学工程研究室,成都610041)2(四川大学电气信息学院医学信息工程学,成都610065)3(广东医学院药理学教研室,湛江524023)4(四川大学华西医院再生医学研究中心心血管疾病研究室,成都610041)摘要:观察体外电刺激对大鼠
2、骨髓间充质干细胞(rBMSCs)向心肌样细胞分化的影响。选用第3、4代rBMSCs用于实验。实验分为3组:(1)电刺激组,rBMSCs分别暴露于1、2、4、6V电压,频率2Hz,脉宽5ms的电刺激,2h/d,连续刺激10d;(2)5-氮胞苷(5-Aza)组,给予10ttmol/L5-Aza诱导24h,然后采用完全培养液培养10d;(3)对照组,采用完全培养液培养10d。通过倒置相差显微镜逐日连续观察细胞的生长状况和形态特征;采用实时荧光定量PCR方法检测GATA4,a一肌动蛋白(a—actin),辅
3、肌动蛋白(ACTN2),心肌肌钙蛋白T(TNNT2)的mRNA表达;采用免疫荧光染色技术检测TNNT2的蛋白表达。结果显示:电刺激组、5-Aza组的GATA4、a—actin、ACTN2和TNNT2mRNA表达与TNNT2蛋白表达明显高于对照组(P4、1—5515.20l301O4EffectsofElectricalStimulationontheDifferentiationofMesenchymalStemCellsintoCardiomyocyte。likeCellsYanZhongqinYangGang2CuiLiao3HeXuelingKuangWeiWuWenchao4LiuXiaojing4LiLiang’1(InstituteofBiomedicalEngineering,WestChinaCollegeofPreclinica5、lMedicineandForensicMedicine,SichuanUniversity,Chengdu610041,China)2(InstituteofMedicalInformaticsEngineering,CollegeofElectricalEngineering&Information。SichuanUniversity,Chengdu610065,China)3(Departmentofpharmacology,GuangdongMedicalCollege,Zhanjiang56、24023。China)4(LaboratoryofCardiovascularDiseases,WestChinaHospital,SichuanUniversity,Chengdu610041,China)Abstract:Theaimofthisstudyistoinvestigatetheeffectsofelectricalstimulation(ES)ontheinductionofratbonemarrowmesenchymalstemcells(rBMSCs)todifferenti7、ateintocardiomyocyte—likecellsinvitro.ThethirdorfourthgenerationoftherBMSCswasrandomlydividedintothreegroups,i.e.ESgroup,5-Azacytidine(5-Aza)group,andcontrolgroup.ThoseintheESgroupwithcompletemediumwereexposedto1,2,4and6V,2Hz,5msESfor2heveryday,lasting8、for10d.Thoseinthe5-Azagroupwereinducedby10p.mol/L5-Azafor24h,thenthemediumwaschangedtocompletemediumwithout5-Aza.Thoseinthecontro1groupwereonlyculturedwithcompletemedium.ThegrowthstatusandmorphologicalfeaturesofrBMSCswereobservedbyinver
4、1—5515.20l301O4EffectsofElectricalStimulationontheDifferentiationofMesenchymalStemCellsintoCardiomyocyte。likeCellsYanZhongqinYangGang2CuiLiao3HeXuelingKuangWeiWuWenchao4LiuXiaojing4LiLiang’1(InstituteofBiomedicalEngineering,WestChinaCollegeofPreclinica
5、lMedicineandForensicMedicine,SichuanUniversity,Chengdu610041,China)2(InstituteofMedicalInformaticsEngineering,CollegeofElectricalEngineering&Information。SichuanUniversity,Chengdu610065,China)3(Departmentofpharmacology,GuangdongMedicalCollege,Zhanjiang5
6、24023。China)4(LaboratoryofCardiovascularDiseases,WestChinaHospital,SichuanUniversity,Chengdu610041,China)Abstract:Theaimofthisstudyistoinvestigatetheeffectsofelectricalstimulation(ES)ontheinductionofratbonemarrowmesenchymalstemcells(rBMSCs)todifferenti
7、ateintocardiomyocyte—likecellsinvitro.ThethirdorfourthgenerationoftherBMSCswasrandomlydividedintothreegroups,i.e.ESgroup,5-Azacytidine(5-Aza)group,andcontrolgroup.ThoseintheESgroupwithcompletemediumwereexposedto1,2,4and6V,2Hz,5msESfor2heveryday,lasting
8、for10d.Thoseinthe5-Azagroupwereinducedby10p.mol/L5-Azafor24h,thenthemediumwaschangedtocompletemediumwithout5-Aza.Thoseinthecontro1groupwereonlyculturedwithcompletemedium.ThegrowthstatusandmorphologicalfeaturesofrBMSCswereobservedbyinver
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