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《無血清體外培養(yǎng)樹突狀細(xì)胞的研究》由會(huì)員上傳分享，免費(fèi)在線閱讀，更多相關(guān)內(nèi)容在教育資源-天天文庫。
1、無血清體外培養(yǎng)樹突狀細(xì)胞的研究作者：張懷東，宋振嵐,李偉平【摘要】本研究的目的是建立對(duì)外周血來源的樹突狀細(xì)胞(dendriticcell,DC)的無血清培養(yǎng)方案。以含胎牛血清(FCS)、人AB血清的培養(yǎng)液作為對(duì)照。分離健康志愿者外周血單個(gè)核細(xì)胞（PBMNC），在不同培養(yǎng)液中分別加入GM-CSF（100ng/ml）、IL-4（500U/ml）培養(yǎng)6天，再分別加入鈣離子載體A23187（100ng/ml）繼續(xù)培養(yǎng)24小時(shí)，于倒置顯微鏡下觀察細(xì)胞形態(tài)，流式細(xì)胞術(shù)分析細(xì)胞表型，MTT比色法檢測(cè)各組DC刺激同種異體外周血
2、T細(xì)胞增殖的能力和DC刺激的T細(xì)胞對(duì)K562細(xì)胞的殺傷作用。結(jié)果表明：無血清培養(yǎng)組培養(yǎng)出典型形態(tài)的DC，其表面CD14分子的表達(dá)明顯減少，CD83、HLA-DR、CDw123分子的表達(dá)明顯增高，具有明顯的刺激同種異體T細(xì)胞增殖的能力和DC刺激的T細(xì)胞對(duì)K562細(xì)胞的殺傷作用。無血清組與兩個(gè)含血清的對(duì)照組比較，組間無顯著性差異（P>0.05）。結(jié)論：無血清培養(yǎng)液培養(yǎng)的DC與含血清的培養(yǎng)液培養(yǎng)的DC相比，無顯著性差異。DC無血清培養(yǎng)具有臨床應(yīng)用前景。【關(guān)鍵詞】樹突狀細(xì)胞；無血清培養(yǎng)液；鈣離子載體；外周血 I
3、nVitroCultivationofDendriticCellswithSerum-free15Medium AbstractThisstudywasaimedtoinvestigatetheprotocolinvitrotoincubatethedendriticcell(DC)derivedfromperipheralbloodmonocytesusingserum-freemediumX-VIVO20.Peripheralbloodmonocytesfromhealthydonorsweretrea
4、tedwith100ng/mlGM-CSFand500U/mlIL-4,respectively.Aftercultivationfor6days,theyweretreatedwith100ng/mlcalciumionophoreA23187.Aftercultivationfor24hoursthecellularmorphologywasobservedunderinvertmicroscope,thesurfacemarkerswereanalyzedbyflowcytometry,theprolif
5、erationofallogeneticTcellswasdetectedbyMTTcolorimetry,thespecificcytotoxicityofTcellsprimedwithDCwasexaminedbyMTTassay.Theresultsshowedthatinallthreegroupswithserum-free,fetalcalfserum(FCS)andhumanABserummediums,cellsdisplayedcharacteristicmorphologicalfeatu
6、resofDC.SimultaneouslyCD14expressionwasdecreased,andCD83,HLA-DRandCDw123expressionwereincreasedonthesecells.Inaddition,DCsculturedwiththesemethodscouldevidentlystimulatetheproliferationofallogeneticTcell.Ascomparedwiththetwocontrolsofserumcontaininggroups,th
7、eculturedcellsintheserum-freegroupsshowedalmostthesameallo-stimulatory15capabilityandcellularmorphologyandsurfacemarkers,andTlymphocytesprimedwiththeculture-derivedDCexhibitedthesimilarkillingactivitytoK562(P>0.05).Itisconcludedthatthereisnosignificancein
8、DCnumbers,morphology,epitopeandabilitytostimulatetheproliferationofallogeneticTcellsbetweenDCinducedbyserum-freeX-VIVO20mediumandDCinducedbyserum-containedmedium.DCculturedandinducedbyserum-free
