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1、ISSN1007-8738細胞與分子免疫學雜志(ChinJCellMolImmunol)2004,20(6)769·論著·文章編號:1007-8738(2004)06-0769-04人胚胎神經(jīng)干細胞向神經(jīng)元分化過程中Notch1基因的表達1122王 飛,李世亭,黃 強,蘭 青(12同濟大學附屬同濟醫(yī)院神經(jīng)外科,上海200065;蘇州大學附屬第二醫(yī)院神經(jīng)外科,江蘇蘇州215004)ExpressionofNotch1geneinthediffer2摘要目的:探討Notch1基因在全反式維甲酸(ATRA)誘導人胚胎神ent
2、iationofthehumanembryonicneural經(jīng)干細胞分化為神經(jīng)元過程中的表達及其發(fā)揮的作用。stemcellstoneurons方法:用不同濃度(0.5、1、5和10μmol/L)的ATRA在體外誘導人胚胎神經(jīng)干細胞,7d后,做NSE免疫熒光染色,計數(shù)分1122WANGFei,LIShi2ting,HUANGQiang,LANQing化為神經(jīng)元的比例。用1μmol/L的ATRA誘導人胚胎神經(jīng)干1DepartmentofNeurosurgery,TongjiHospitalofTongjiUnivers
3、ity,Shang2細胞,于誘導前、誘導3d和誘導7d,提取細胞的總RNA,用2hai200065;DepartmentofNeurosurgery,theSecondAffiliatedHospital半定量RT2PCR法檢測Notch1基因的表達。結果:與對照組ofSuzhouUniversity,Suzhou215004,China相比較,ATRA能顯著提高人胚胎神經(jīng)干細胞分化為神經(jīng)元的比例(P<0.01)。其中1μmol/L的ATRA誘導人胚胎神經(jīng)Abstract干細胞分化為神經(jīng)元的比例最高,為(29.20±1
4、.09)%。AIM:ToexploretheexpressionandroleofNotch1geneintheNotch1基因在ATRA誘導后表達明顯下調(diào)(P<0.001)。結論:differentiationofthehumanembryonicneuralstemcellstoneu2ATRA能顯著提高人胚胎神經(jīng)干細胞分化為神經(jīng)元的比例,ronsinducedbyall2trans2retinoic2acid(ATRA).METHODS:Notch1基因在人胚胎神經(jīng)干細胞分化為神經(jīng)元的過程中表達Thehumane
5、mbryonicneuralstemcellswereinducedwithvari2量下調(diào)。ousconcentrationsofATRA(0.5,1,5and10μmol/L)invitro.關鍵詞:神經(jīng)干細胞;Notch1基因;表達;分化Immunofluorescencestainingwasperformedaftersevendays,中圖分類號:Q254 文獻標識碼:AandneuronswerequantifiedbycountingNSEpositivecells.Thenhumanembryon
6、icneuralstemcellswereinducedtodiffer2 近年來,人類神經(jīng)干細胞的分離、培養(yǎng)成功,為entiatewith1μmol/LATRA.TotalRNAwasextractedfromthecellsbeforeinducing,inducedfor3days,andfor7days.The神經(jīng)發(fā)育的研究和中樞神經(jīng)系統(tǒng)疾病的治療提供了expressionofNotch1genewastestedbysemi2quantitativeRT2一條新的思路。無論是作為腦組織移植的供體,還PCR.
7、RESULTS:Theratioofneuralstemcellstodifferentiate是作為體內(nèi)誘導分化的靶細胞,神經(jīng)干細胞(neutraltoneuronswasimprovedgreatlybyATRAinductionascom2stemcell,NSC)都為中樞神經(jīng)系統(tǒng)的功能重建和神經(jīng)paredwithcontrolgroup(P<0.01).1μmol/LATRAwasthe再生提供了光明的應用前景。大腦內(nèi)神經(jīng)組織存在optimaldoseforinductionofthehumanembryoni
8、cneuralstem多種細胞類型,包括神經(jīng)元、星形膠質(zhì)細胞及少突膠celldifferentiation.Thepercentageofneuronsinducedby質(zhì)細胞等。大腦的功能主要依賴于神經(jīng)元并通過神1μmol/LATRAwas(29.20±1.09)%.Theexpressionof經(jīng)信號傳導的方式來實現(xiàn)。因此,定
