【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 CL 的关键更多还原

【Abstract】 Objective: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更多还原