Biomaterial Database

Improvement in the efficiency of the in vitro transformation assay method using BALB/3T3 A31-1-1 cells. 1995

T Tsuchiya, and M Umeda

Safety Evaluation Center, Central Research Laboratory, Chiba, Japan.

In order to improve the in vitro transformation assay using BALB/3T3 cells, which is routinely conducted with minimal essential medium (MEM) containing 10% fetal calf serum (FCS), we examined the effect of a new medium after the cells had been treated with carcinogens. Preliminary experiments suggested that the use of T medium (modified DME/F-12) supplemented with insulin, transferrin, ethanolamine and sodium selenite plus 2% FCS resulted in a high transformation frequency. The present study confirmed that this medium was very efficient at inducing transformation foci. Transformation frequency was highest when 12-O-tetradecanoylphorbol-13-acetate (TPA) was started from 1 week after the carcinogen treatment. According to the new protocol using this medium, the transformation frequency was five times higher and transformation foci appeared much earlier than with the protocol using usual MEM plus 10% FCS medium. In addition, we tested several typical carcinogens and a promoter in order to confirm the applicability of the new protocol to the two-stage transformation assay. N-Methyl-N'-nitro-N-nitro-soguanidine (MNNG) and benzo[a]pyrene, but not noncarcinogenic benzo[e]pyrene, induced transformation foci. In the metabolic activation system dimethylnitrosamine gave a highly positive transformation. Okadaic acid, a non-TPA-type tumor promoter, enhanced transformation of MNNG-initiated cells. These studies demonstrate that the use of the proposed medium drastically improves transformation frequency in the two-stage in vitro transformation assay.

UI	MeSH Term	Description	Entries
D008769	Methylnitronitrosoguanidine	A nitrosoguanidine derivative with potent mutagenic and carcinogenic properties.	Methylnitrosonitroguanidine,Nitrosomethylnitroguanidine,Nitrosonitromethylguanidine,MNNG,N-Methyl-N'-nitro-N-nitrosoguanidine,N Methyl N' nitro N nitrosoguanidine
D002471	Cell Transformation, Neoplastic	Cell changes manifested by escape from control mechanisms, increased growth potential, alterations in the cell surface, karyotypic abnormalities, morphological and biochemical deviations from the norm, and other attributes conferring the ability to invade, metastasize, and kill.	Neoplastic Transformation, Cell,Neoplastic Cell Transformation,Transformation, Neoplastic Cell,Tumorigenic Transformation,Cell Neoplastic Transformation,Cell Neoplastic Transformations,Cell Transformations, Neoplastic,Neoplastic Cell Transformations,Neoplastic Transformations, Cell,Transformation, Cell Neoplastic,Transformation, Tumorigenic,Transformations, Cell Neoplastic,Transformations, Neoplastic Cell,Transformations, Tumorigenic,Tumorigenic Transformations
D003470	Culture Media	Any liquid or solid preparation made specifically for the growth, storage, or transport of microorganisms or other types of cells. The variety of media that exist allow for the culturing of specific microorganisms and cell types, such as differential media, selective media, test media, and defined media. Solid media consist of liquid media that have been solidified with an agent such as AGAR or GELATIN.	Media, Culture
D004128	Dimethylnitrosamine	A nitrosamine derivative with alkylating, carcinogenic, and mutagenic properties. It causes serious liver damage and is a hepatocarcinogen in rodents.	Nitrosodimethylamine,N-Nitrosodimethylamine,NDMA Nitrosodimethylamine,N Nitrosodimethylamine,Nitrosodimethylamine, NDMA
D000818	Animals	Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA.	Animal,Metazoa,Animalia
D013755	Tetradecanoylphorbol Acetate	A phorbol ester found in CROTON OIL with very effective tumor promoting activity. It stimulates the synthesis of both DNA and RNA.	Phorbol Myristate Acetate,12-Myristoyl-13-acetylphorbol,12-O-Tetradecanoyl Phorbol 13-Acetate,Tetradecanoylphorbol Acetate, 4a alpha-Isomer,12 Myristoyl 13 acetylphorbol,12 O Tetradecanoyl Phorbol 13 Acetate,13-Acetate, 12-O-Tetradecanoyl Phorbol,Acetate, Phorbol Myristate,Acetate, Tetradecanoylphorbol,Myristate Acetate, Phorbol,Phorbol 13-Acetate, 12-O-Tetradecanoyl,Tetradecanoylphorbol Acetate, 4a alpha Isomer
D016475	3T3 Cells	Cell lines whose original growing procedure consisted being transferred (T) every 3 days and plated at 300,000 cells per plate (J Cell Biol 17:299-313, 1963). Lines have been developed using several different strains of mice. Tissues are usually fibroblasts derived from mouse embryos but other types and sources have been developed as well. The 3T3 lines are valuable in vitro host systems for oncogenic virus transformation studies, since 3T3 cells possess a high sensitivity to CONTACT INHIBITION.	3T3 Cell,Cell, 3T3,Cells, 3T3
D051379	Mice	The common name for the genus Mus.	Mice, House,Mus,Mus musculus,Mice, Laboratory,Mouse,Mouse, House,Mouse, Laboratory,Mouse, Swiss,Mus domesticus,Mus musculus domesticus,Swiss Mice,House Mice,House Mouse,Laboratory Mice,Laboratory Mouse,Mice, Swiss,Swiss Mouse,domesticus, Mus musculus