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1、小鼠胚胎干细胞诱导为甲状腺细胞的实验研究【摘要】目的 探讨体外定向诱导胚胎干细胞(ESCs)分化为甲状腺细胞的可行性。方法 E14小鼠ESCs诱导发育为拟胚体，再逐步添加促甲状腺素（TSH）、胰岛素（insulin）、KI等共培养。观察细胞分化过程的形态学变化，并以体外培养的成年小鼠甲状腺细胞为阳性对照；RT-PCR法检测甲状腺细胞分化相关基因TSHR、PAX8、NIS、TPO、Tg等表达情况；激光共聚焦显微镜双色荧光检测甲状腺细胞分化标记物PAX8和TTF-1的共表达;在荧光检测得甲状腺细胞分化率的基础上，送检电镜观察分化细胞的超微结构。结果诱导
2、培养第6d分化细胞中有甲状腺细胞特有基因PAX8、NIS、TPO、Tg、TSHR的表达；第8d检测到分化细胞中甲状腺细胞标记物TSHR、TTF-1、PAX8、TTF-2的表达；细胞形态类似对照甲状腺细胞。结论 ESCs经胚胎体发育阶段，在特定条件下可定向分化为甲状腺细胞。【关键词】干细胞；分化；甲状腺；拟胚体Experimentalstudyondirecteddifferentiationofembryonicstemcellsintothyrocyte-likecellsinvitro.ZHANGHong,JIANGNing-yi,LIUShe
3、ng,etal.(NuclearMedicineDepartment,theSecondAffiliatedHospitalofSunYat-senUniversity,Guangzhou510120,Guangdong,P.R.China)Abstract:ObjectiveToinvestigatethefeasibilityofdirecteddifferentiationofembryonicstemcells(ESCs)intothyrocyte-likecellsinvitro.MethodsMurineE14ESCswerecultu
4、redinmethylcellulosesemisolidmediumtoformembryoidbodies(EBs).TheseEBsweretransferredforfurtherinductiveculturewiththestepwiseadditionofgrowthfactors-TSH,insulinandKIintotheculturemedium.Duringdifferentiation,cellmorphologywasobservedthroughphasecontrastmicroscopyandcomparedwit
5、hthenormalthyroidcellsfrommouse.Themolecularmarkersofthyroidcellswereperformedbyindirectimmunofluorescsentanalysisunderfluorescsentmicroscopy.GeneexpressionsofthyroidspecificmRNAwereanalyzedbyreversetranscription-polymerasechainreaction(RT-PCR)formoleculesTSHR,PAX8,NIS,TPOandT
6、g.ResultsAfterEBsformation,onday6offurthercultureaddedwithinductivefactorsTSH,insulinandKI,ESCs-derivedcellsexpressedthyroid-specificgenessuchasPAX8,NIS,TPO,TgandTSHR.Ontheeighthday,theseESCs-derivedthyrocyte-likecellsexpressedTSHR,TTF-1,PAX8andTTF-2.Morphologyofthesemarkerspo
7、sitivedifferentiatedcellswassimilartothosenormalthyroidcells.ConclusionESCscandifferentiateintothyrocyte-likecellsundergiveninductiveconditionsandrelatedgrowthfactorsinvitro.Keywords:Stemcells;Differentiation;Thyroidgland;Embryoidbody近年来，甲状腺疾病的发生率逐年升高，各种治疗方法如甲状腺肿瘤的手术，有些涉及到甲状腺全
8、切除或次全切除，导致术后甲状腺功能减退；甲状腺机能亢进症的放射性核素131I治疗疗效确切，但其最大的不足是出现甲状腺功能减
