人脱落乳牙干细胞骨向分化相关分子Runx2的动态表达王立媛1，2，刘大勇1，贾智11天津医科大学口腔医院，天津市300070，2天津市口腔医院，南开大学口腔医院，天津市300041DynamicexpressionofRunx2geneprofileduringosteogenesisinstemcellsfromhumanexfoliateddeciduousteethWangLi-yuan1,2,LiuDa-yong1,激aZhi11DentalHospital,Tian激nMedicalUniversity,Tian激n300070,China;2Tian激nStomatologicalHospital,StomatologicalHospitalofNankaiUniversity,Tian激n300041,China摘要背景：Runx2被认为是成骨基因表达的主要调节因子，是一种向成骨细胞分化的不可或缺的转移因子，在成骨细胞发育、分化、调控、骨钙化形成及骨修复过程中起着极其重要的作用。目的：观察人脱落乳牙干细胞生物学特征，探讨乳牙干细胞骨向分化潜能，以及不同时间点成骨相关转录因子Runx2的动态表达规律。方法：体外分离、培养人脱落乳牙干细胞。流式细胞仪检测乳牙干细胞表面标志。体外脂向诱导分化4周后进行油红O染色，检测脂滴形成情况；矿化诱导21d进行茜素红染色，检测矿化结节形成情况。于成骨诱导的不同时间点，用RT-PCR技术检测Runx2的动态表达。结果与结论：通过酶消化法和有限稀释法获得了乳牙干细胞，流式细胞仪检测显示CD146和STRO-1均有不同程度的表达。成脂诱导油红O染色可见橙红色阳性颗粒。矿化诱导茜素红染色呈阳性。RT-PCR技术检测Runx2在成骨诱导第0天已有表达，0-6d成明显增强趋势，6-12d显著下降，12-18d表达再次上升，以后相对平稳。结果表明乳牙干细胞可体外分离、培养，表达间充质干细胞表面标志，具有成脂和成骨向分化能力。Runx2在乳牙干细胞成骨分化各阶段均有表达，早期和晚期表达上升，中期表达有下降趋势。中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程全文链接：关键词：组织构建；骨组织工程；人脱落乳牙干细胞；Runx2；骨向分化；动态表达；Abstract：BACKGROUND:Runx2isconsideredtothemainregulatoryfactorofosteogenicgeneexpressionandbenecessaryforosteoblastdifferentiation,itplaysanextremelyimportantroleintheosteoblastdevelopment,differentiation,regulation,bonecalcificationformationandbonerepair.OBJECTIVE:Toobservethebiologicalpropertiesofmesenchymalstemcellsfromhumanexfoliateddeciduousteeth,exploretheosteogenicdifferentiationpotentialofdeciduousteethstemcells,andobservethedynamicexpressionofRunx2geneatvaryingtimepoints.---本文于网络，仅供参考，勿照抄，如有侵权请联系删除---METHODS:Thestemcellsfromhumanexfoliateddeciduousteethwereisolatedandculturedinvitro.Thecellsurfaceantigenwasdetectedwithflowcytometry.Thethirdpassagecellswereculturedintheadipogenicmediumfor4weeks,andoilredOstainingwasconductedtotestlipiddropletsformation.Thethirdpassagecellswereculturedintheosteogenicmediumfor21days,andmineralizednodulesweredetectedbyalizarinredstaining.Runx2mRNAdynamicexpressionwasdetectedwithsemi-quantitativeRT-PCRatdifferenttimepoints.RESULTSANDCONCLUSION:Thestemcellsfromhumanexfoliateddeciduousteethwereobtainedbyenzymedigestionandlimiteddilutionmethods.Flowcytometryresultsshowedthat,CD146andSTRO-1wereexpressedtovaryingdegrees.OilredOstainingrevealedsalmonpinkpositiveps.Alizarinredstainingshowedpositiveexpression.RT-PCRresultsshowedthat,Runx2expressionwasfoundatday0,up-regulatedfromday0today6,andsubsequentlydroppedwithanexpressionbottomatday12,afterthatasecondexpressionpeakoccurredatday18,followedbyastablyregulation.Thestemcellsfromhumanexfoliateddeciduousteethcanbeisolatedandculturedinvitro,expresssurfaceantigenofmesenchymalstemcells,andhavethepotentialsofdifferentiatingintoadipocytesandostetoblasts.Runx2geneprofilesaredy...