Brodeur B R, Tsang P, Larose Y
J Immunol Methods. 1984 Jul 6;71(2):265-72. doi: 10.1016/0022-1759(84)90073-5.
Hybridoma cells injected intraperitoneally into mice induce formation of ascites tumours and production of ascites fluid containing high levels of monoclonal antibody. Several parameters affecting the growth of the immunoglobulin-producing tumours have been studied in order to define optimal conditions for ascitic fluid formation and monoclonal antibody production. Using hybridomas produced by fusing SP2/0 myeloma cells with immunized mouse spleen cells we have shown: (1) that the optimal number of hybridoma cells required to induce an ascites tumour was between 6 and 32 X 10(5) cells; (2) that each mouse should be treated with a maximum of 0.5 ml of pristane; (3) that the priming period for pristane should be 14 days prior to the injection of cells; (4) that ascites formation and monoclonal antibody production is significantly better in males; and finally (5) that the age of mice used should range between 43 and 78 days. Under these conditions each mouse produces on average 7-10 ml of ascites fluid, containing a high level of antibody, over a maximum period of 6 days. The animals should start producing between the 5th and 9th day and usually survive 11-16 days after being injected with the tumour cells.
将杂交瘤细胞腹腔注射到小鼠体内可诱导腹水肿瘤的形成,并产生含有高水平单克隆抗体的腹水。为了确定腹水形成和单克隆抗体产生的最佳条件,研究了几个影响产生免疫球蛋白肿瘤生长的参数。利用SP2/0骨髓瘤细胞与免疫小鼠脾细胞融合产生的杂交瘤,我们发现:(1)诱导腹水肿瘤所需的最佳杂交瘤细胞数量在6至32×10⁵个细胞之间;(2)每只小鼠最多应注射0.5 ml的降植烷;(3)降植烷的预处理期应在注射细胞前14天;(4)雄性小鼠的腹水形成和单克隆抗体产生明显更好;最后(5)所用小鼠的年龄应在43至78天之间。在这些条件下,每只小鼠在最长6天的时间内平均产生7 - 10 ml含有高水平抗体的腹水。动物应在第5至9天开始产生腹水,注射肿瘤细胞后通常存活11 - 16天。