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《體外誘導(dǎo)大鼠骨髓間充質(zhì)干細(xì)胞向神經(jīng)細(xì)胞分化的實(shí)驗(yàn)研究》由會(huì)員上傳分享，免費(fèi)在線閱讀，更多相關(guān)內(nèi)容在工程資料-天天文庫(kù)。
1、大連醫(yī)科大學(xué)學(xué)報(bào)2014年第36卷第1期(JournalofDalianMedicalUniversity2014．Vol．36No．1)7基礎(chǔ)醫(yī)學(xué)doi:10．11724/jdmu．2014．01．02體外誘導(dǎo)大鼠骨髓間充質(zhì)干細(xì)胞向神經(jīng)細(xì)胞分化的實(shí)驗(yàn)研究李雙月，王哲敏，陳若霖，戚媛，劉爽，樸豐源(大連醫(yī)科大學(xué)勞動(dòng)衛(wèi)生與環(huán)境衛(wèi)生學(xué)教研室，遼寧大連116044)［摘要］目的對(duì)大鼠骨髓間充質(zhì)干細(xì)胞(bonemarrowmesenchymalstemcells，BMSCs)進(jìn)行分離、培養(yǎng)與鑒定，并探討全反式維甲酸(retinoicacid，
2、ＲA)、堿性成纖維細(xì)胞生長(zhǎng)因子(fibroblastgrowthfactor，basic，bFGF)和表皮生長(zhǎng)因子(epidermalgrowthfactor，EGF)聯(lián)合誘導(dǎo)BMSCs分化為神經(jīng)細(xì)胞的可行性。方法全骨髓貼壁法分離培養(yǎng)BMSCs，觀察細(xì)胞形態(tài)及生長(zhǎng)增殖情況;流式鑒定細(xì)胞表面標(biāo)志物CD29、CD34、CD90;選用第3代細(xì)胞，經(jīng)ＲA、bF-GF和EGF聯(lián)合誘導(dǎo)后，細(xì)胞免疫化學(xué)染色檢測(cè)神經(jīng)細(xì)胞標(biāo)志物神經(jīng)元特異性烯醇化酶(neuronspecificenolasen，NSE)的表達(dá)。結(jié)果體外培養(yǎng)的BMSCs呈成纖維細(xì)胞樣，第
3、3、4、5代BMSCs的生長(zhǎng)曲線均呈S形，活性無(wú)明顯差異。BMSCs的均一性較好，第3代細(xì)胞CD29、CD90陽(yáng)性率均在90%以上，而CD34陽(yáng)性率僅為0．58%;BMSCs經(jīng)誘導(dǎo)后分化為神經(jīng)細(xì)胞，并表達(dá)神經(jīng)細(xì)胞標(biāo)志NSE。結(jié)論成功建立BMSCs的體外培養(yǎng)體系，所得細(xì)胞純度高、生物學(xué)特征穩(wěn)定，并可誘導(dǎo)分化為神經(jīng)細(xì)胞，為移植治療神經(jīng)系統(tǒng)損傷提供實(shí)驗(yàn)基礎(chǔ)。［關(guān)鍵詞］骨髓間充質(zhì)干細(xì)胞;神經(jīng)細(xì)胞;分化;誘導(dǎo);培養(yǎng)［中圖分類號(hào)］Ｒ322．2［文獻(xiàn)標(biāo)志碼］A文章編號(hào):1671－7295(2014)01－0007－04［引用本文］李雙月，王哲敏，陳
4、若霖，等．體外誘導(dǎo)大鼠骨髓間充質(zhì)干細(xì)胞向神經(jīng)細(xì)胞分化的實(shí)驗(yàn)研究［J］．大連醫(yī)科大學(xué)學(xué)報(bào)，2014，36(1):7－10．Neuron－likecellsdifferentiatedfromratmesenchymalstemcellsinvitroLIShuang－yue，WANGZhe－min，CHENＲuo－lin，QI－Yuan，LIUShuang，PIAOFeng－yuan(DepartmentofOccupationalandEnvironmentalHealth，DalianMedicalUniversity，Dalian
5、116044，China)［Abstract］ObjectiveToisolateandcultureratbonemarrowmesenchymalstemcells(BMSCs)andinducethemdifferen-tiatetoneuralcells．MethodsBMSCswereisolatedfromratsbywallstickingmethod．Thenthecellswereidentifiedwithmorphology，proliferationandthesurfacemarkers．Neuron－lik
6、ecellsfromBMSCswereinducedbythecombinationofret-inoicacid(ＲA)，fibroblastgrowthfactor，basic(bFGF)andepidermalgrowthfactor(EGF)，andtheirneuronspecificeno-lasen(NSE)weredetectedbyNSEimmunohistochemistry．ＲesultsCulturedBMSCslikefibroblast－shapedinvitro．thGrowthcurvesofthe3r
7、d，4and5th－generationBMSCsandtheiractivitieswerenotsignificantlydifferent．FCMdetectedthatCD29andCD90werepositivelyexpressed，butCD34wasnegativelyexpressed．TheinducedBMSCsshowedneuron－likemorphologyandexpressedNSE，theneuronmarker．ConclusionWehaveestablishedanefficientandst
8、eadymethodtoobtainBMSCs．BMSCscoulddifferentiatedintoneuronswhichwasanidealsourceforclinicalcelltransplantation
