Orikasa M, Takasugi K, Fujiwara A, Motoda S, Shimizu F
J Immunol Methods. 1985 Dec 17;85(1):75-85. doi: 10.1016/0022-1759(85)90275-3.
A simple and convenient method for efficiently establishing 8-azaguanine-resistant mutant leukemia and myeloma cell lines (for example, the T cell lines Jurkat and CCRF-CEM, human myeloid/macrophage-like cell lines HL60 and U937, Burkitt lymphoma line Raji and the human myeloma line RPMI 8226), is described. The method relies on culturing the cell lines in RPMI 1640 medium containing 8-azaguanine and supplemented with 15% heat-inactivated fetal calf serum and large amounts of amino acids and vitamins, and removes the necessity for pretreatment with mutagenic reagents such as ethyl methylsulfonate or X-irradiation. The possibility of obtaining mutant cell lines using the method described here is about 15 times greater than using media without high levels of amino acids and vitamins. Hybridomas produced between mitogen-activated human peripheral blood lymphocytes and an 8-azaguanine-resistant Jurkat mutant cell line (established by this method) were shown to produce soluble T cell-derived macrophage activating factor (MAF)-like material.
本文描述了一种简单便捷的方法，可高效建立对8-氮杂鸟嘌呤具有抗性的突变白血病和骨髓瘤细胞系（例如T细胞系Jurkat和CCRF-CEM、人髓样/巨噬细胞样细胞系HL60和U937、伯基特淋巴瘤细胞系Raji以及人骨髓瘤细胞系RPMI 8226）。该方法是将细胞系培养于含有8-氮杂鸟嘌呤的RPMI 1640培养基中，并添加15%热灭活胎牛血清以及大量氨基酸和维生素，从而无需使用如甲基磺酸乙酯或X射线照射等诱变剂进行预处理。使用本文所述方法获得突变细胞系的可能性比使用不含高水平氨基酸和维生素的培养基高出约15倍。有丝分裂原激活的人外周血淋巴细胞与一株对8-氮杂鸟嘌呤具有抗性的Jurkat突变细胞系（通过该方法建立）杂交产生的杂交瘤，被证明可产生可溶性T细胞衍生的巨噬细胞激活因子（MAF）样物质。
