新乡医学院学报JournalofXinxiangMedicalCollegeVo1．24No．6Nov．2007·541··论著·改良组织块贴壁法培养大鼠主动脉平滑肌细胞及鉴定王倩，舒茂琴，江明宏，李建涛，康振峻(1．第三军医大学附属西南医院心血管内科，重庆400038；2．解放军第252医院心血管内科，河北保定071000)摘要：目的以简捷、高效的方法获取原代和传代的大鼠血管平滑肌细胞(VSMC)，为进一步的科学研究提供实验材料。方法采用改良的组织块贴壁法培养原代大鼠VSMC，2．5g·L胰蛋白酶消化传代，并用相差显微镜、电镜、荧光显微镜及激光共聚焦显微镜对细胞进行鉴定。结果95％的组织块接种成活，相差显微镜下细胞呈典型的“峰一谷”状生长，胞体长梭形；电镜下可见粗、细肌丝沿细胞长轴平行排列；荧光显微镜及激光共聚焦显微镜检测一肌动蛋白可见细胞胞浆内呈阳性显色，细胞核不显色。结论本方法培养VSMC简便、易操作，获得的细胞纯度和成活率较高。关键词：大鼠；血管平滑肌细胞；细胞培养；主动脉中图分类号：Q813．11文献标识码：A文章编号：1004-7239(2007)06—0541—04Cultureandidentificationofratsvascularsmoothmusclecellswithmodifiedtissue-pieceinoculationWANGQian，SHUMao·qin，JIANGMing·hong，etal(1．DepartmentofCardiology，SouthwestAffiliatedHospitaloftheThirdMilitaryMedicalUniversity，Chongqing400038，China；2．DepartmentofCardiology，the252ndHospitalofPeopleSLiberationA，Hebei071000，China)Abstract：ObjectiveToobtainvascularsmoothmusclecellsoforiginalgenerationandpassageofratinsimpleandel-fectivewayfortherelatedresearches．MethodsTheoriginalgenerationcellswereobtainedbymodifiedtissue—pieceinoculationandwith0．25％trypsindigestiontoobtainpassagecells．Thecellswereidentifiedbycontrastphasemicroscope，electronmicro—scope，fluorescencemicroscopeandconfocalmicroscopy．Results95％inoculatedtissuepiecesweresurvived．Undercontrastphasemicroscope，cellsshowedthetypical“peak—valley”morphological，cellbodieswerefusiformshape．Underelectronmicro—scopethickmyofilamentandthinfilamentrangedparallellyalongcellmacroaxis．Underfluorescencemicroscopeandconfoealmi，croscopy．endochylemaof一actinwasbeenseenthatitwasstainedpositive．Andthenuceluswasntbeenstained．ConclusionItis...
