Bohbot A, Uttwiller F, Fujita R, Mousson B, Fabre M, Oberling F, Poindron P
Université Louis Pasteur (ULP), Centre de Recherches d'Immunologie et Hématologie, Strasbourg, France.
In Vitro Cell Dev Biol Anim. 1993 May;29A(5):362-70. doi: 10.1007/BF02633983.
Despite more than 50 attempts and the use of various methods, it has been impossible to establish homologous hybridomas between human mature macrophages and 8-azaguanine-resistant U-937 clones prepared in the laboratory. To rule out the possibility that these clones were unsuitable for the selection of hybrids, a study of their properties was done. It was shown that U-937 wild type cells were able to produce HPRT, whereas 8-azaguanine (8-aza)-resistant clones did not. Curiously, exonic and intronic HPRT sequences were undetectable both in wild type and in 8-aza-resistant cell genomes, under conditions where they were detected in control cells. Chromosome analysis of the clone UM9, one of the most frequently used in fusion experiments, revealed many qualitative and quantitative differences with the U-937 wild type cells. 8-aza-resistant U-937 cells were capable of fusion with human macrophages and gave rise to heterokaryons and probably to synkaryons, which survived for weeks without dividing in hypoxanthine-aminopterin-thymidine medium. The results could be interpreted in terms of the existence of a transacting negative regulatory mechanism of the macrophage genome on the proliferative capacity of homospecific hybridomas.
尽管进行了50多次尝试并使用了各种方法,但仍无法在人类成熟巨噬细胞与实验室制备的8-氮杂鸟嘌呤抗性U-937克隆之间建立同源杂交瘤。为了排除这些克隆不适合用于杂交选择的可能性,对它们的特性进行了研究。结果表明,U-937野生型细胞能够产生次黄嘌呤磷酸核糖转移酶(HPRT),而8-氮杂鸟嘌呤(8-aza)抗性克隆则不能。奇怪的是,在对照细胞中能检测到外显子和内含子HPRT序列的条件下,野生型和8-aza抗性细胞基因组中均未检测到这些序列。对融合实验中最常用的克隆UM9进行染色体分析,发现其与U-937野生型细胞存在许多定性和定量差异。8-aza抗性U-937细胞能够与人类巨噬细胞融合,形成异核体,可能还形成了合核体,这些合核体在次黄嘌呤-氨基蝶呤-胸腺嘧啶核苷培养基中存活数周而不分裂。这些结果可以解释为巨噬细胞基因组对同种特异性杂交瘤增殖能力存在反式作用的负调控机制。