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《大鼠原代肝細(xì)胞、星狀細(xì)胞、枯否細(xì)胞和肝竇內(nèi)皮細(xì)胞的同步分離與培養(yǎng)-論文.pdf》由會(huì)員上傳分享，免費(fèi)在線閱讀，更多相關(guān)內(nèi)容在行業(yè)資料-天天文庫(kù)。
1、·532·doi10．3969~．issn．1673—4254．2014．04．20JSouthMedUniv,2014，34(4)：532—537基礎(chǔ)研究大鼠原代肝細(xì)胞、星狀細(xì)胞、枯否細(xì)胞和肝竇內(nèi)皮細(xì)胞的同步分離與培養(yǎng)李洋，蔡雙明，張莉莉，李旭南方醫(yī)科大學(xué)南方醫(yī)院急診科，廣東廣州510515摘要：目的探索和建立同步分離大鼠原代肝細(xì)胞、星狀細(xì)胞、枯否細(xì)胞和肝竇內(nèi)皮細(xì)胞，并進(jìn)一步鑒定和培養(yǎng)的方法。方法聯(lián)合應(yīng)用原位灌注、離體灌注、梯度密度離心和差速貼壁等分離方法，獲取4種大鼠肝內(nèi)原代細(xì)胞，采用形態(tài)學(xué)觀察、免疫熒光染色和墨汁吞噬實(shí)驗(yàn)等方法對(duì)所分離
2、的各種原代細(xì)胞進(jìn)行純度鑒定。結(jié)果通過原位灌注結(jié)合離體灌注、密度梯度離心結(jié)合差速貼壁等分離手段，成功實(shí)現(xiàn)了大鼠原代肝細(xì)胞、星狀細(xì)胞、枯否細(xì)胞以及肝竇內(nèi)皮細(xì)胞的同步分離，且所得原代細(xì)胞得率、活力及純度等指標(biāo)均符合后續(xù)細(xì)胞實(shí)驗(yàn)要求。結(jié)論通過簡(jiǎn)易方法同步分離大鼠原代肝細(xì)胞、星狀細(xì)胞、枯否細(xì)胞以及肝竇內(nèi)皮細(xì)胞是可行的。關(guān)鍵詞：原代培養(yǎng)；肝細(xì)胞；星狀細(xì)胞；枯否細(xì)胞；肝竇內(nèi)皮細(xì)胞；原位灌注；密度梯度離心SimultaneousisolationandprimarycultureofrathepatocytesfhepaticstellatecellslK
3、upffer’ScellsandhepaticsinusendothelialcellsLIYang,CAIShuangmin~ZHANGLili,LIXuDepartmentofEmergencyMedicine,Na咖ngHospital，SouthernMedicalUniversity,Guangzhou510515,ChinaAbstract：ObjectiveToestablishamethodforsimultaneousisolationandprimarycultureofrathepatocytes,hepaticste
4、llatecells,Kupffer’Scellsandhepaticsinusendothelialcells．MethodsBycombining協(xié)situperfusion,invitroperfusion，densitygradientcentrifugationanddifferentialadhension,primaryrathepatocytes,hepaticstellatecells,Kupffer’Scellsandhepaticsinusendothelialcellswereobtained．Thepurityof
5、thesecellswereassessedwithmorphologicalobservation,immunofluorescentstainingandinkphagocytosisassa~ResultsWesuccessfullyobtainedthe4primarycellssimultaneouslybycombininginsituperfusionwithinvitroperfusion,densitygradientcentrifugation,anddifferentialattachment．Thecellyield
6、rate，cellviabilityandpurityallmetrequirementsforthesubsequentcellexperiment．ConclusionThecombinedcellisolationandculturemethodisfeasibletoisolateprimaryrathepatocytes，hepaticstellatecells，Kupffer’Scellsandhepaticsinusendothelialcellssimultaneously．Keywords：primaryculture；h
7、epatocytes；hepaticstellatecells；Kupffer’Scells；hepaticsinusendothelialcells；insituperfusion；densitygradientcentrifugation肝臟的主要細(xì)胞構(gòu)成為上皮細(xì)胞(肝細(xì)胞、膽管上胞分離手段，通過一次分離獲取多種肝內(nèi)原代細(xì)胞皮細(xì)胞)及問質(zhì)細(xì)胞(肝竇內(nèi)皮細(xì)胞、枯否細(xì)胞、星狀細(xì)具有重要意義。筆者參閱國(guó)內(nèi)外文獻(xiàn)，結(jié)合多次試驗(yàn)摸胞、間皮細(xì)胞、血管平滑肌細(xì)胞、隱窩細(xì)胞、神經(jīng)細(xì)胞索，總結(jié)出一套實(shí)用、穩(wěn)定而高效的分離方法，歸納報(bào)道等)?。目前的研究熱
8、點(diǎn)集中于肝細(xì)胞、肝竇內(nèi)皮細(xì)胞、如下。枯否細(xì)胞和星狀細(xì)胞。在大鼠體內(nèi)，上述4種細(xì)胞各自的原代分離、鑒定和培養(yǎng)方法已有大量文獻(xiàn)報(bào)道。近1材料和方法年來，隨著人們對(duì)肝臟生理病理機(jī)制研究
