成牛血清低分子量成分在杂交瘤细胞体外“无血清”培养中的利用

THE USE OF LOW-MOLECULAR-WEIGHT COMPONENTS FROM ADULT CATTLE SERUM IN IN VITRO CULTURE OF HYBRIDOMA CELLS

利用适量的(3.5—14%)成牛血清低分子成分超滤液(LMW-CS,排阻M=300 00)、10mg/L转铁蛋白(T)、10 mg/L胰岛素(Ⅰ)、20μ mol/L乙醇胺(E)、40 n mol/L硒酸钠(S,当以RPMT1640为基础液时用)等诸补充成分替代胎牛血清加到基础液RPMI1640或1:1(v/v)的IMDM与Ham's F_(12)混合液中构成本实验性'无血清'培养基。培养在此种培养基中的杂交瘤细胞其最高细胞密度可以达到甚至超过培养在含胎牛血清的生长培养基中的最高细胞密度。对于杂交癌细胞生长,4%的LMW-CS基本满足要求,T、I、E、S四种补充成分中乙醇胺(E)是首位影响因子;LMW-CS与TIES两部分之间的协同作用极为显著,两者缺一不可。

This paper reported a 'serum-free' medium supporting the growth of hybridoma cells in vitro.Low molecular weight fraction from adult cattle serum (LMW-CS) passed throuth an Amicon PM30 filter(M. W. exclusion=30000), 10mg/L transferrin(T), 10mg/L insulin (Ⅰ), 20μmol/L ethanolamine (E), and 40 nmol/L sodium selenite(S)as the replacement of fetal calf serum (FCS) in culture medium were added to basal medium, RPMl1640 or 1:1 mixture (by volum) of IMDM (Iscove's Modified Dulbecco's Medium) and Ham's F12, to compose the experimental 'serum-free' medium. Maximum density of hybridoma cells cultured in such medium can reach and sometimes exceed that in FCS-containing medium. For the growth of hybridoma cells, 4% LMW-CS in the medium was found to be able to meet the basic requirement; ethanolamine (E) may be the primary growth-supporting factor among the 4 essential supplements of T, I, E, and S; and a significant synergistic action between LMW-CS and TIES on the cell growth was found.