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《大鼠肝細(xì)胞與Kupffer細(xì)胞共同培養(yǎng)對(duì)兩者功能和形態(tài)影響.doc》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、大鼠肝細(xì)胞與Kupffer細(xì)胞共同培養(yǎng)對(duì)兩者功能和形態(tài)影響作者：寇晨光曹成波燕曉雯羅兵周巖冰【摘?！慷挠^察大鼠肝實(shí)質(zhì)細(xì)胞與Kupffer細(xì)胞體外共同培養(yǎng)吋對(duì)兩者生長(zhǎng)、形態(tài)及功能的影響。方法釆用原位二步IV型膠原灌注法＞Percol1液密度梯度離心法分離Wistar大鼠肝實(shí)質(zhì)細(xì)胞與Kupffer細(xì)胞;體外將肝實(shí)質(zhì)細(xì)胞與Kupffer細(xì)胞按6:1比例共同培養(yǎng)。觀察不同情況下肝細(xì)胞生存時(shí)間和形態(tài)，每隔24h檢測(cè)培養(yǎng)上清液屮清蛋白和谷丙轉(zhuǎn)氨酶(ALT).谷草轉(zhuǎn)氨酶(AST)的水平，并在培養(yǎng)36h用放免法檢測(cè)上清液中口細(xì)胞介素1(IL1)、口細(xì)胞介素6(IL6)、腫瘤壞死
2、因子a(TNFa)含量。結(jié)果單獨(dú)培養(yǎng)組肝細(xì)胞的生長(zhǎng)、增殖迅速，并向止常肝細(xì)胞的形態(tài)演變，肝細(xì)胞可培養(yǎng)存活至15d;共同培養(yǎng)組肝細(xì)胞生長(zhǎng)增殖緩慢，細(xì)胞可培養(yǎng)存活至10do共同培養(yǎng)組上清液中清蛋白水平在24、36、48、60h比單獨(dú)培養(yǎng)組低(t二2.551?3.139,P<0.05);ALT.AST水平在24、36、48、60h比單獨(dú)培養(yǎng)組高(t=2.446-3.108,P<0.05);共同培養(yǎng)組Kupffer細(xì)胞保持IL1、IL6、TNFa分泌功能，而單獨(dú)培養(yǎng)組未檢測(cè)到IL1、IL6、TNFao結(jié)論大鼠Kupffer細(xì)胞和肝實(shí)質(zhì)細(xì)胞在適宜的培養(yǎng)條件下可進(jìn)
3、行共同培養(yǎng)，二者可以保持良好的分泌功能，可用于實(shí)驗(yàn)研究?！娟P(guān)鍵詞】肝細(xì)胞;枯否細(xì)胞;共同培養(yǎng)技術(shù);清蛋白類;細(xì)胞因子類;大鼠[ABSTRACT]ObjectiveToobservethegrowth,morphology,andfunctionofhepatocyteswhenculturedinvitrowithKupffercelIsinrats?MethodsTnsitucollagenasetwostepperfusionmethodandlowspeedgradientcentrifugtionbypercollfluidwereusedtoisolat
4、eparenchymalhepaticcells(PHC)andKupffercel1sinWistarrat,respective!y.CocultivationofPHCandKupfferce11swasdoneinvitroaccordingto6:lratio.Thelivetimeandformofthecel1sunderdifferentcircumstanceswereobserved?ALTandASTlevelsinculturesupernatantweredetectedat24hourinterval,andtheconcentrati
5、onofIL1,IL6andTNFameasuredbyradioininiunoassayat36hours?ResultsInsolitaryculturegroup,thehepatocytesgrewquicklyanddevelopedtotheformofnormallivercells,whichcouldsurvivel5days;forthoseincoculturegroup,thecel1sgrewandgeneratedslowly,whichsurvivedlOdays.Thelevelsofalbumeninsupernateofcoc
6、ulturegroup,at24h,36h,48hand60h,werelowerthanthatinsolitaryculturegroup(t=2.551-3.139,P<0.05),andthatofALTandAST,atthesametimepointsasabove,werehigher(t=2.446一3?108,P&It;0.05)?Incoculturegroup,thesecretoryfunctionofIL1,IL6,andTNFawasretained,andinsolitaryculturegroup,noTL1,IL6,andT
7、NFaweredetected?ConclusionHepatocytesandKupffercel1sofratcanbeculturedtogetherunderasuitablecondition,andthefavourablesecretoryfunctionofthecellsbemaintainedforempiricalstudy?[KEYWORDS]hepatocyte;Kupffercell;coculturetechnique;albumins;cytokines;rats肝細(xì)胞分離、培養(yǎng)是研究肝臟和肝細(xì)胞的良好方法。盡管肝細(xì)胞在體內(nèi)擁有相當(dāng)
8、強(qiáng)的增殖能
