长寿老人永生细胞库的建立与保存

Establishment and preservation of immortal lymphoblastoid cell lines of long-lived people

目的 建立长寿老人永生细胞库 ,为长寿遗传机制的研究提供永久的研究材料。方法 用 EB病毒 (Epstein- Barr virus)转染长寿老人外周血 B淋巴细胞 ,采用 2 0 % FBS RPMI1 640培养液 ,配合环胞霉素 A和 PHA,置 37℃ 5% CO2 培养箱内培养。及时加液或少量换液 ,当细胞增长到一定数量时分瓶传代 ,分瓶传代到 1 6瓶及以上予以冻存 ,之后复苏以证明冻存成功情况。结果 成功地对广西巴马长寿乡壮族长寿家庭的 2 4个个体建立了永生细胞株。其中 ,百岁老人 6株 ,90～ 99岁老人 1 2株 ,长寿老人≤ 66岁的直系亲属 6株。结论 所有建成的细胞株冻存后复苏的成功率为 1 0 0 %。经染色体 G显带分析未见异常 ,表明建立细胞株的方法稳定。

Objective To provide perpetual research materials for longevity studies, we established immortal lymphoblastoid cell lines of long lived people.Methods The B lymphoid cells,which were isolated from the long lived people's blood samples,were transformed by Epstein Barr Virus and cultured in 20% FBS RPMI1640 cultivation medium with Cyclosporine A,PHA etc,in incubator of 37 ℃ and 5% CO 2.Added or changed the cultivation medium in time.After the number of cells increased to abounding amount, they were transferred to another culture bottle. When they were cultured up to 16 or more bottles,they were storaged in liquid nitrogen.The immortal lymphoblastoid cell lines were verified successfully by revivification.Results 24 immortal lymphoblastoid cell lines of long lived people of Bama were triumphally obtained,including 6 long lived people,12 nonagenarians and 6 healthy individuals younger than 66 years old.Conclusions All of the immortal lymphoblastoid cell lines were successfully revivificated after been frozen in liquid nitrogen.By the analysis of chromosome Gimesa banding , no abnormity was found.The approach to establish immortal lymphoblastoid cell lines is reliable.