建立巴马长寿老人经EBV转染永生细胞株的方法  

ESTABLISHMENT OF THE METHOD OF OBTAINING IMMORTAL CELL LINES BY EPSTEIN-BARR VIRUS FROM LONGEVOUS PEOPLES IN BAMA GUANGXI

作　　者：吕泽平[1] 郑陈光[1] 杨泽[2] 吴建洲[1] 韦勇[1] 龙桂芳[3] 潘尚领[3] 史晓红[2] 孙玉茹[2] 胡才友[1] 许代娣[1] 庞国防[1] 孙逸平[2] 陈建超 

机构地区：[1]广西壮族自治区江滨医院,南宁530021 [2]卫生部老年医学研究所遗传学研究室,北京100730 [3]广西医科大学,南宁530021 [4]广西巴马县长寿研究所,巴马547500

出　　处：《广西医科大学学报》2002年第5期624-626,共3页Journal of Guangxi Medical University

基　　金：广西自然科学基金项目 (桂科自 9912 0 3 2 )资助

摘　　要：目的 :建立用 EBV转化长寿人群外周血淋巴细胞永生细胞株 (L CL)的方法。方法 :用 EBV转染 8份百岁以上健康老人 ,2 0份 89～ 99岁健康老人及 8份 6 6岁以下健康人的血样共 36份。采用 2 0 % FBS RPMI16 4 0培养液 ,配合环孢菌素 A、植物血凝素 (PHA) ,置 37℃ 5 % CO2 培养箱内培养。结果 :成功地获得 2 4份永生细胞株 (其中 ,百岁老人 6份 ,89～ 99岁老人 12份 ,6 6岁以下 6份 )。结论 :采血后 2 4～ 4 8h之间接种 ,转化成功率高。转染后 2～ 3周 ,大部分标本出现细胞分裂、变形 ,标志着转化成功。超过 5周仍无细胞分裂者即发生细胞凋亡。PHA有缓解细胞凋亡的作用 ,但不能使细胞分裂。污染是引起转染细胞株凋亡的主要因素。严格的无菌操作、保持培养细胞的适当密度、适时换液、打散较大的细胞克隆团是培养 L CLObjective:To establish the method for line by way of peripheral B lymphoid cell that was transformed by Epstein Barr Virus Method:The lymphoid cell were isolated from the blood samples including 8 centenarians, 20 nonagenarians and 8 healthy individuals of less than 66 years old in Bama, Guangxi B lymphoid cell was transformed by Epstein Barr Virus and cultured in 20% FBS RPMI1640 cultivation medium with cyclosporine A?PHA etc incubated in 37℃ Result:24 immortal longevity cell lines were obtained successfully, which included 6 centenarians, 12 nonagenarians and 6 healthy individuals of less than 66 years old Conclusion:After blood sample was collected with in 24~48 hours,there was more possibility of success transforming Epstein Barr Virus into the target cell In general,in living cell transformed in 2~3 weeks cell division and distortion occured It means that the transformation is successful PHA may reduce protosis of the cultivated cells, but could not induce the transformation and division of the cells Otherwise, over 5 weeks, the final was cell protosis It should be noted that to prevent from pollution, keep moderate cell density, change cultivate medium in time and break up clone cell mass are very important

关 键 词：EB病毒 长寿老人 细胞培养 巴马 广西 永生细胞株 

分 类 号：Q813.11[生物学—生物工程]