HGPRT缺陷型T淋巴瘤细胞系的建立与鉴定  

Establishment and Identification of HGPRT Deficient T Cell Line

作　　者：翁秀芳[1] 吴雄文[1] 梁智辉[1] 陆盛军[1] 卢小玲[1] 张彩娥[1] 钟茂华[1] 陈雪玲[1] 龚非力[1] 

机构地区：[1]华中科技大学同济医学院基础医学院免疫学系,武汉430030

出　　处：《华中科技大学学报（医学版）》2006年第6期701-704,共4页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金重大项目资助(No.30490241);国家"973"计划资助项目(No.2001CB510008)

摘　　要：目的通过突变诱导次黄嘌呤鸟嘌呤磷酸核糖转移酶(HGPRT)缺陷型的T淋巴瘤细胞系,为制备T细胞杂交瘤提供利于筛选的亲代细胞。方法通过乙基甲磺酸对Jurkat细胞进行突变,逐步提高培养液中6-巯基鸟嘌呤(6-TG)的浓度,筛选出对6-TG具有稳定抗性的单克隆细胞株Jur16-TG细胞。比较Jurkat细胞与Jur16-TG细胞在含6-TG培养液和HAT培养液中的生长情况。结果在含20μg/ml 6-TG的培养液中,Jur16-TG细胞能够稳定生长,而Jurkat细胞在培养1周后基本死亡。在HAT培养液中氨基喋呤的浓度为4×10-8mol/L或8×10-8mol/L时,Jur16-TG细胞4 d内基本死亡,而Jurkat细胞至少能存活7~10 d。Jur16-TG细胞与正常外周血淋巴细胞(PBL)的融合实验显示,35%的Jur16-TG细胞与PBL融合。结论突变筛选出的单克隆细胞株Jur16-TG具有6-TG抗性和在HAT培养液中不能生长的特性,证实该细胞为HGPRT缺陷型细胞株,并且该细胞株可有效地与原代淋巴细胞融合,从而为T细胞杂交瘤的制备提供了利于筛选的亲代永生化的T细胞。Objective To establish a HGPRT deficient T cell line from Jurkat cell line for T cell hybridization.Methods The Jurkat cells were cultured with EMS and selected by increasing concentration of 6-TG.The TG-resistant mutant Jur1^(6-TG) cells were cultured in the medium containing 6-TG or HAT.Results Jur1^(6-TG) cells could survive in medium containing 20 μg/ml 6-TG,while Jurkat cells died within 1 week.In medium containing HAT with aminopterin concentration of 4×10^(-8) mol/L and 8×10^(-8) mol/L,Jur1^(6-TG) cells died rapidly within 4 days,whereas Jurkat cells survived at least 7-10 days.And 35% Jur1^(6-TG) cells successfully fused with the PBLs. Conclusion The TG-resistant and aminopterin-sensitive character of Jur1^(6-TG) reveals that the Jur1^(6-TG) cell is deficient in HGPRT.Together with the efficient fusion with PBLs,the Jur1^(6-TG) will be a good parent cell line for preparation of T cell hybridoma.

关 键 词：次黄嘌呤鸟嘌呤磷酸核糖转移酶缺陷 6-巯基鸟嘌呤抗性 次黄嘌呤/氨基喋呤/胸腺嘧啶(HAT)培养液 T细胞杂交瘤 

分 类 号：R392.12[医药卫生—免疫学]