用于骨髓瘤细胞和杂交瘤细胞增殖以及杂交瘤细胞抗体产生的无血清培养基  

PROLIFERTION OF MYELOMA CELL AND HYBRIDOMA CELL AND MONOCLONAL ANTIBODY PRODUCTION OF HYBRIDOMA CELL IN SERUM-FREE MEDIUM

作　　者：周维松[1] 蔡少华[1] 

机构地区：[1]中国农业科学院生物技术研究中心,北京100081

出　　处：《中国农业科学》1992年第3期82-88,共7页Scientia Agricultura Sinica

摘　　要：小鼠骨髓瘤SP_2/O细胞和小鼠杂交瘤7E8细胞不需经低浓度血清的适应性培养即能在无血清培养基Ⅰ(SFM—Ⅰ)中良好生长、增殖。培养获得的最大活细胞密度分别为2.57×10~6/ml和1.88×10~6/ml,对大于1×10~6/ml活细胞密度的维持时间均为84小时。这两方面均优于用无血清培养基Ⅱ(SFM—Ⅱ)或用有血清培养基Ⅱ(SCM—Ⅱ)培养的效果。SFM—Ⅰ培养7E8细胞的培养上清中,特异性单克隆抗体ELISA效价为1∶800,与SFM—Ⅱ和SCM—Ⅱ培养上清的效价相同,SFM—Ⅰ可望用作杂交瘤细胞体外大规模无血清培养生产单克隆抗体的首选培养基。 SFM—Ⅱ培养的细胞从初始密度增殖至1×10~6/ml时,其增殖速度与有血清培养基相近,大于SFM—Ⅰ培养细胞的增殖速度。用SFM—Ⅱ进行SP_2/O、7E8和大鼠骨髓瘤IR983细胞的连续传代培养,细胞始终圆整、立体感强,具有高度活力和较快的增殖速度。SFM—Ⅱ适宜用于骨髓瘤和杂交瘤细胞的无血清传代培养。Mouse myeloma SP_2/0 cell and hybridoma 7E8 cell, without adaptation in low serum concentration medium, can be well cultivated in serum-free medium I (SFM-I) . The maximun viable cell density of 2.57×10~6 cells/ml in the case of SP_2/0 cell and 1.88×10~6 cells/mlin the case of 7E8 cell were attained. An equal of 84hrsmaitaining time of the viable cell density above 1×10~6 cells / ml was achieved when either SP_2 / 0 cell or 7E8 cell was cultured in SFM-I . These data were better than those achieved in SFM-Ⅱ and in serum-contianing medium Ⅱ (SCM-Ⅱ) . The same valence of 1:800 ELISA titer of monoclonal antibody in culture supernatant was reached when 7E8 cell was cultured in SFM-Ⅰ, SFM-Ⅱ and SCM-Ⅱ respectively. It suggests that there is a trail to use SFM-Ⅰ for large-scale serum-free culture of hybridoma cell.As the cell density going up to 1×10~6 cells/ml from the initial cell density in cell proliferation test, a similar proliferation rate of 7E8 cell and SP_2/ 0 cell in either SFM-Ⅱ or SCM-Ⅱ was acheived, which was higher than that in SFM-I. As whown in SFM-Ⅱ contin ual subculturing test, SP_2/0, 7E8 and rat myeloma IR983 cells were always round, stereoscopic, high viable and fast prolific SFM-Ⅱ is suggested as the medium of choice for myeloma and hybridoma cell subculturing.

关 键 词：抗体 骨髓瘤 杂交瘤 无血清培养基 

分 类 号：S857.4[农业科学—临床兽医学]