title: Establishment of an abalone digestive gland cell line secreting various glycosidases in protein-free culture.
abstract: A cell line designated as ADG was established from an abalone digestive gland using ERDF medium supplemented with 8% fetal bovine serum (FBS), 8% abalone hemolymph, and high concentrations of NaCl, KCl, MgCl(2), MgSO(4), and CaCl(2). ADG cells proliferated better in protein-free medium than in FBS-supplemented medium. Among 9 kinds of media examined, ERDF medium was shown to be optimal for cell growth. ADG cells secreted 13 different kinds of glycosidases in protein-free medium: α-L-fucosidase, β-L-fucosidase, α-D-galactosidase, β-D-galactosidase, N-acetyl-α-D-galactosaminidase, N-acetyl-β-D-galactosaminidase, α-D-glucosidase, β-D-glucosidase, N-acetyl-α-D-glucosaminidase, N-acetyl-β-D-glucosaminidase, α-D-mannosidase, β-D-mannosidase, β-D-xylosidase, and 1-3 xylanase. When ADG cells were cultured in Grace's insect cell medium, the activity of some secreted glycosidases increased 25-fold to 65-fold per cell as compared with control cells cultured in ERDF medium. ADG - abalone digestive gland; ERDF - enriched RDF; FBS - fetal bovine serum; L-15 - Leibovitz's L-15 media; DME - Dulbecco's modified Eagle medium; F-12 - nutrient mixture (Ham); LDF - L-15; DME: F-12 = 10 : 7 : 3; MEM - minimum essential medium; RPMI - RPMI medium 1640; 199 - media 199; GIC - Grace's insect cell medium; pNP -p -nitrophenol.
authors.affiliation: Cellular Regulation Technology Laboratory, Graduate School of Genetic Resources Technology, Kyushu University, Hakozaki, Higashiku, Fukuoka, 812, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Kusumoto
authors.forename: S
authors.initials: S
authors.lastname: Shirahata
authors.forename: Y
authors.initials: Y
authors.lastname: Katakuta
authors.forename: H
authors.initials: H
authors.lastname: Murakami
authors.forename: Y
authors.initials: Y
authors.lastname: Kamei