Jahn S, Grunow R, Kiessig S T, Specht U, Matthes H, Hiepe F, Hlinak A, Von Baehr R
Institute for Medical Immunology, Humboldt-University, Berlin, G.D.R.
J Immunol Methods. 1988 Feb 24;107(1):59-66. doi: 10.1016/0022-1759(88)90009-9.
50 fusion experiments were carried out to analyse heterohybridization efficiencies on mouse myeloma cells of the P3 X63 Ag8/653 line with human lymphocytes derived from peripheral blood, bone marrow, lymph node, spleen or synovial fluid. We found higher yields of growing and human Ig-producing hybridoma lines when lymphocytes from spleen or lymph node were fused. Although primary hybridomas could be established from fusions with bone marrow-derived cells, only in nine out of 1616 initially seeded wells was Ig production registered. Four fusions using immune cells from synovial fluid were made without success. Independently of the source of lymphocytes pokeweed mitogen (PWM) prestimulation had no enhancing effect on the percentage of wells with cell growth and this did not alter the IgM:IgG ratio in primary hybridomas (9:1), although cells from all compartments used here (with the exception of bone marrow cells) could be stimulated with PWM to produce both IgG and IgM in cultures. Cryopreserved lymphocytes from different sources could be used for fusions with comparable results registered for the fresh material.
进行了50次融合实验，以分析P3 X63 Ag8/653系小鼠骨髓瘤细胞与源自外周血、骨髓、淋巴结、脾脏或滑液的人淋巴细胞的异种杂交效率。我们发现，当脾脏或淋巴结中的淋巴细胞融合时，生长中的和产生人Ig的杂交瘤细胞系产量更高。虽然可以通过与骨髓来源的细胞融合建立初代杂交瘤，但在最初接种的1616个孔中，只有9个孔检测到Ig产生。使用滑液免疫细胞进行的4次融合未成功。无论淋巴细胞来源如何，商陆有丝分裂原（PWM）预刺激对细胞生长孔的百分比没有增强作用，并且这不会改变初代杂交瘤中的IgM:IgG比例（9:1），尽管此处使用的所有区室的细胞（骨髓细胞除外）都可以用PWM刺激，在培养物中产生IgG和IgM。来自不同来源的冷冻保存淋巴细胞可用于融合，新鲜材料的结果与之相当。
