资源描述：
《体外大鼠肝卵圆细胞的长期培养方法.doc》由会员上传分享，免费在线阅读，更多相关内容在工程资料-天天文库。
1、体外大鼠肝卵圆细胞的长期培养方法尹寿新1,周震S马美雪3,杨景波J羊东晔II中南大学湘雅二医院消化内科，湖南省长沙市410011；2长沙市第一医院消化内科，湖南省长沙市410005；3徐州市中心医院消化内科，江苏省徐州市221009Long・te「mcultureofhepaticovalcellsinratsinvitroYinShou-xin',ZhouZhen2,MaMei-xue3,YangJing・bo[YangDong・ye〔'DepartmentofGastroenterology&Hepatology,theSecon
2、dXiangyaHospitalofCentralSouthUniversity,Changsha410011,HunanProvince,China;2DepartmentofGastroenterology&Hepatology,theFirstChangshaHospital,Changsha410005,HunanProvince,China;'DepartmentofGastroenterology&Hepatology,XuzhouCentralHospital,Xuzhou221009,JiangsuProvince,C
3、hina摘要背景：肝卵圆细胞是目前公认的成体肝干/祁细胞，但其体外长期培养时会不可避免地丢火干细胞的活性。目的：探索大鼠肝卵関细胞体外氏期培养的方法。方法：构建2•乙酰胺基易7部分肝切除肝再工大鼠模型，通过酶消化和Percoll梯度离心分离纯化大鼠异质卵圆细胞并进行免疫染色鉴定，用含表皮生长因了、白血病抑制因了的培养皋体外长期培养，后撤去表皮生长因了、□血病抑制因子，通过形态学的观察和分了标志物的检测判断苴能否保持〒/祖细胞活性。结果与结论：采用含表皮生长因了、白血病抑制因了的培养基体外培养卵圆细胞4个月后，大鼠异质卵圆细胞仍能表达刖
4、•细胞标志物ALB、胆管上皮细胞标志物CK-19,经不倉表皮⑴长因子、门血病抑制因了的培养液继续培养后，卵圆细胞胎肝标志AFP表达量迅速下降。该研究结果表明大鼠肝卵圆细胞在表皮生长因了、n血病抑制因了等培养条件卜可长期增殖并保持干细胞活性。中国纽织工程研究杂志出版内容巫点：纽织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程全文链接：关键词：组织构建；组织工程；卵圆细胞；长期培养；表皮生长因子；白血病抑制因子；国家口然科学基金;Abstract:BACKGROUND:Hepaticovalcellsare
5、recognizedasstem/progenitorcellscurrently，however,Iong-termcultureofhepaticovalcellscaninevitablyresultinthelossofcellactivity.OBJECTIVE:ToexploreaIong-termculturemethodofhepaticovalcellsinvitro.METHODS:Partiallyhepatectomizedratmodelwasestablishedbyusing2AAF/PH.Therege
6、Deratedliverwasdigestedbycollagenase,andhepaticovalcellswereisolatedandpurifiedbydensitygradientcentrifugationandidentifiedbyimmunocytochemistry.Hepaticovalcellswereculturedinamediumcontainingepidermalgrowthfactorandleukaemiainhibitoryfactor.Thenepidermalgrowthfactorand
7、leukaemiainhibitoryfactorwereremovedafterseveralmonths,anditsabilityofmaintainingstem/progenitorcellactivitieswasdeterminedbasedonmorphologyandmolecularmarkers・RESULTSANDCONCLUSION:BothhepatocytemarkerALBandbiliaryepithelialcellmarkerCK・19werefoundafterhepaticovalcellsw
8、ereculturedinamediumcontainingepidermalgrowthfactorandleukaemiainhibitoryfactorfor4months・Whileintheabseneeofe
