The use of low-molecular-weight components from adult cattle serum in culture of hybridoma cells in vitro.
Adequate amounts (3.5-14%) of low-molecular weight fractions from adult cattle serum (LMW-CS) passed through an /Amicon PM30 filter (mol. wt. exclusion = 30000), 20 microM ethanolamine (E), 10 mg/l transferrin (T), 10 mg/l insulin (I), and 40 nM sodium selenite (S), used as the substitutes for fetal calf serum (FCS), were added to the basic medium, RPMI1640 or a 1:1 mixture (by volume) of IMDM (Iscove's Modified Dufbecco'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 was the primary growth-supporting factor among the 4 essential supplements of T, I, E, and S; and a significant synergistic action on the cell growth was found between LMW-CS and TIES.
Du
S Y
SY
Institute of Applied Microbiology, Heilongjiang Academy of Sciences, Harbin.
D000818:Animals; D001781:Blood Component Removal; D002417:Cattle; D002452:Cell Count; D002455:Cell Division; D016895:Culture Media, Serum-Free; D006825:Hybridomas / Q000166:cytology*; D051379:Mice; D008970:Molecular Weight
D016895:Culture Media, Serum-Free