构建原代分离培养与鉴定小鼠肺泡ⅱ型上皮细胞的方法及模型

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1、万方数据中国组织工程研究与临床康复第74誊第15嬲2010—04—09出版JournalofClinicalRehabilitativeTissueEngineeringResearchApril9，2010VoL14，No．15构建原代分离培养与鉴定小鼠肺泡II型上皮细胞的方法及模型：l：：

2、：☆郑金旭，黄振杰，汤艳，丁明《辎％槲№《目螭∞赫Ⅻ目目∞#∞黼《$目黼黼熵m$#Ⅻ∞∞Ⅻ∞《《Ⅻ∞《踹∞蛳Ⅻ％㈣㈣@∞Ⅻ№Ⅻ#∞∞械№#镕剿№@黼Ⅻ《黼ⅫⅫ№∞∞蛹∞∞#@ⅫⅫmW㈥∞W酣∞≈$∞Ⅻ∞酬％Ⅻ№@@Ⅻ∞滞∞《№‰∞@姆Ⅻ㈣№№Ⅻ∞Ⅻ$Ⅻ∞㈣Ⅻ㈣ⅫⅫ∞∞∞∞《

3、ⅫWWN嘲镕㈣㈣Ⅻ斓鲫《Ⅻ@∞《瓣Ⅻ㈣∞∞《磷№*‰猕Isolation，primarycultureandidentificationoftype11alveolarepithelialcellsfrommiceZhengJin-xu，HuangZhen_jie，TangYan，DingMingAbstractBACKGROUND：TypeIIalveolarepithelialcells(AECII)playanimportantroleinlungdevelopmentandregulationof．1ungfunction．1tiscloselyrelat

4、edwiththegenesisanddevelopmentofpulmonaryfibrosis．1ungcancerandsoon．1nordertomakean『仃vitrostudyofthesediseases．astableandeffectivecelImodeIforisolation．primarycultureandidentificationofAECIIshouldbeestablished．OBJECTlVE：TOdevelopareliablemethodfortheisolation，purification，primarycultu

5、reandidentificationofAECIIfrOmmice，andprovideafoundationforthefurtherstudyofpulmonaryfibrosisandlungcancer．METHODS：①Thelungtissueofmicewasdigestedwithlow-concentrationtrypsinandcollagenaseI．Thatis，lungtissuesofmicewerecutintosmallpieces，digestedwithtrypsin，andterminatedthedigestionbya

6、ddingDMEMcontainingfetalbovineserum．Thecellsweredigestedonceagainusingtrypsin．andaddingcollagenaseIaftertermination．(函AEC11waspurifiedbydifferentialcentrifugationandimmuneadherenceforprimaryculture．ThepneumocytessuspensionwasincubatedinIgGcultureplate，andliquidcontainingnon-adherentce

7、llswereonceagainculturedinIgGcultureplate，thensupernatantwereabandoned，andthecellswereculturedjnculturedishandcultureplate．ThemorphologyandgrowthcharacteristicsofprimarilyculturedAEC11wereobservedbyanjnvertedmicroscope；theyield．purityandviabilityofAEC11weredetermined．AEC11wasidentifie

8、dbyimmunocytochemicalstainingsudactantapoproteinA(SP-A)andSP-C，andthespecificcelIstructurewasobservedunderatransmissionelectronmicroscope．RESULTSANDCONCLUSlON：Bythismethod。anamountof(4．8+1．2)x105AEC11wereharvestedfromeachmousewithapurityof(85土2．4)％andaviabilityof(92±2．4)％．Underaninver

9、tedmi