【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。更多还原

【Abstract】 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.更多还原