Clone M-3

Origin:  mouse F1 (CxDBA), clone from melanoma Cloudman S91.

            Science 1966. 154: 1186.

Morphology:  epithelial-like

Mode of cultivation: monolayer

Conditions for cultivation: medium - F10 or F12

serum -  FBS 10% (F12) or HS 15%/FBS 2.5% (F10)

subculture procedure - cells detach from flask using trypsin 0.25%:

EDTA  0.02% (1:3), split ratio 1:4 - 1:8, optimal population density 2.0-3.0104 cells/cm2

cryoconservation - growth medium, 10% DMSO, 2.0-3.0106 cells/ml in ampule

Viability after cryoconservation:     94% (0 passage, dye trypan blue)

Sterility: tests for bacteria, fungi and mycoplasma were negative

Species: karyological and isoenzymological (LDH, G6PD) analysis

Karyology:   2n= 40, variability in the range between 76-86 chromosomes,

modal number of chromosomes 83, number of markers - 2 (routine dye),

some cells have microchromosomes (ATCC).

Plating efficiency:    less than 1% (ATCC)

Tumorigenicity:  tumorigenic in syngeneic animals

virus susceptibility: herpes simplex, vaccinia, pseudorabies, vesicular stomatitis (Indiana).

Melanin production for at least 33 passages

Applications:  virology, tumorigenicity, cell biology.

Collections: ATCC CCL 53.1; ECACC 87081806; SPBIC.