Habituation in suspension-cultured soybean cells to thiamine and its precursors

When thiamine concentration in subculture medium was rapidlylowered to nil, soybean cells in suspension became necroticand stopped growing entirely. When it was gradually lowered,cell growth was vigorous until the concentration was reducedto 7.8?10^–3 mg/liter. The cells at this level of thiamineceased growing for a time, but prolonged culture in the samemedium resulted in the appearance of fresh white cells whichcould be easily distinguished from the old brown, necrotic cellsin the aggregates. These new cell lines could be subculturedwith further reduction in the thiamine supply, growing as largeraggregates of about 4 mm in diameter. New cell lines were similarly obtained by prolonged culturesin media containing a thiamine precursor; three lines appearedto be habituated to the pyrimidine moiety and one to the thiazolemoiety. The latter cell line could be subcultured without thiamineand its precursors for at least eight passages. These habituatedcells were characterized by the increase of the dry to freshweight ratio and by their growth in large aggregates. ¹Present address: Section of Phytochemical Research, Eisai Co.,Ltd., Kawashima, Gifu 483, Japan. (Received December 15, 1978; )