CONDITIONED MEDIUM FROM MACROPHAGE CELL-LINES SUPPORTS THE SINGLE-CELL GROWTH OF HYBRIDOMAS
CONDITIONED MEDIUM FROM MACROPHAGE CELL-LINES SUPPORTS THE SINGLE-CELL GROWTH OF HYBRIDOMAS
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DOI：
10.1089/hyb.1986.5.255
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发表时间：
1986-09-01
期刊：
影响因子：
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通讯作者：
GECZY, CL
中科院分区：
文献类型：
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作者：
RATHJEN, DA;GECZY, CL
本研究的目的是建立是否条件培养基（CM）从巨噬细胞系将支持杂交瘤的生长条件下常用的杂交实验和克隆抗原特异性杂交瘤。将来自巨噬细胞系J774、WEHI 274、WEHI 265和PU 5的CM在克隆期间支持单细胞生长的能力与来自驻留小鼠腹膜细胞、EL 4小鼠胸腺瘤细胞、L929小鼠纤维肉瘤和驻留腹膜细胞饲养层的培养物的CM进行比较。来自J774、L929和驻留腹膜细胞的CM支持与巨噬细胞饲养层相同水平的单细胞生长。J774和L929 CM在最终浓度为25%时最有效，新鲜培养基补充有20%胎牛血清（FCS）。J774 CM支持杂交瘤生长的能力通过预先用LPS刺激而不是PMA刺激而增加。与用巨噬细胞饲养层获得的杂交瘤相比，融合实验中使用的来自LPS刺激的J774细胞的CM导致杂交瘤数量增加。
The aim of this study was to establish whether conditioned medium (CM) from macrophage cell lines would support the growth of hybridomas under conditions commonly used in hybridization experiments and in cloning of antigen-specific hybridomas. The ability of CM from macrophage cell lines J774, WEHI 274, WEHI 265, and PU 5 to support single-cell growth during cloning was compared with CM from cultures of resident mouse peritoneal cells, EL 4 mouse thyoma cells, L929 mouse fibrosarcoma, and feeder layers of resident peritoneal cells. CM from J774, L929, and resident peritoneal cells supported single-cell growth at the same level as the macrophage feeder layer. J774 and L929 CM were most effective at a final concentration of 25% with fresh medium supplemented with 20% fetal calf serum (FCS). The ability of J774 CM to support hybridoma growth was increased by prior stimulation with LPS but not PMA. CM from LPS-stimulated J774 cells used in fusion experiments resulted in increased numbers of hybridomas compared with those obtained with macrophage feeder layers.