小鼠脾臟來(lái)源樹突狀細(xì)胞的體外擴(kuò)增培養(yǎng)

《小鼠脾臟來(lái)源樹突狀細(xì)胞的體外擴(kuò)增培養(yǎng)》由會(huì)員上傳分享，免費(fèi)在線閱讀，更多相關(guān)內(nèi)容在學(xué)術(shù)論文-天天文庫(kù)。

1、小鼠脾臟來(lái)源樹突狀細(xì)胞的體外擴(kuò)增培養(yǎng)作者：黎輝，曹宇皎，黃軍華，劉俊峰【摘要】目的：對(duì)體外誘導(dǎo)小鼠脾臟單個(gè)核細(xì)胞分化成的樹突狀細(xì)胞(DC)的生物學(xué)表型及功能進(jìn)行檢測(cè)，并與骨髓來(lái)源的DC進(jìn)行比較。方法：分離小鼠脾臟單個(gè)核細(xì)胞，在體外培養(yǎng)的條件下通過(guò)添加25ng·ml-1的粒單核細(xì)胞集落刺激因子和25ng·ml-1的白細(xì)胞介素4(IL4)將其誘導(dǎo)為DC，分別從細(xì)胞形態(tài)、表型以及功能3個(gè)方面對(duì)其進(jìn)行檢測(cè)，并與骨髓來(lái)源的DC進(jìn)行比較。結(jié)果：小鼠脾臟來(lái)源的DC經(jīng)磷酸脂多糖(LPS)刺激成熟后其表面顯示出明顯的樹枝狀突起，高表達(dá)CD11c、CD86及MH

2、CⅡ類分子，并具有極強(qiáng)的刺激同種異基因淋巴細(xì)胞增殖的能力，其特征與骨髓來(lái)源的DC相差無(wú)幾。結(jié)論：小鼠脾臟單個(gè)核細(xì)胞能夠被誘導(dǎo)為具有正常形態(tài)、表型、功能的DC，為DC的功能研究以及進(jìn)一步制備相應(yīng)的腫瘤疫苗提供又一可靠來(lái)源。【關(guān)鍵詞】脾臟;骨髓;樹突狀細(xì)胞;誘導(dǎo);小鼠　Dendriticcells(DC)arethemostpotentantigenpresentingcellsthatareresponsibleforprimingnativeTcellsintheimmuneresponse.ImmunotherapyofDC/tumorvacc

3、inehasbecomeanimportanttherapeuticstrategyagainsttumors[12].However,thebasicandclinicalapplicationsofDCshavebeen11limitedbythelowyieldandpurityofDCsgeneratedbytraditionalbonemarrowderivation.Inthisstudy,weinducedmousespleenmononuclearcellsintodendriticcellsinvitro,andthencomp

4、aredthebiologicalcharacteristicsofthemwiththoseofbonemarrowmononuclearcellsderivedDCs.　　1Materialsandmethods　　1.1Materials　　Mainreagents:recombinantmousegranulocytemacrophagecolonystimulatingfactor(GMCSF),recombinantmouseinterleukin4(IL4),recombinantmouseleukemiainhibitor

5、yfactor(LIF,Peprotech),lipopolysaccharide(LPS,Sigma);mitomycinC(MMC)andmethabenzthiazuron(MTT,Duchfo).DCsculturemedium:highglucoseDMEMplus15%fetalcalfserumplus25mg·L-1rmGMCSFandrmIL4.Mice:sixtoeightweeksoldfemaleBALB/cand129micewerepurchasedfromAnimalCenterofGuiyangMedicalCo

6、llege.　　1.2Methods11　　1.2.1InductionofDCfrommousespleenCellswereflushedoutofspleenof129mice.Mononuclearcellsattheinterfacewerecollectedbycentrifugationoveralympholytegradient.Theywerewashedandthenculturedin6wellplatefor2hwiththedensityof5×104ml-1.Thesuspendedcellswereremoveda

7、ndtheadherentcellswereculturedwithDCculturemedium.LPSwasaddedonday7andthesuspendedcellswereharvested24hlater,whileatthesametimemousebonemononuclearcellswereinducedintoDCs.　　1.2.2MorphologyobservationCellswereobservedundermicroscopedailyandsuspendedcellswereidentifiedbyelectron

8、microscopy.　　1.2.3CellsurfacemarkeranalysisCellswerecollected