Origin: mouse C3H, smooth muscle-like cells from brain tumor induced in vivo by ethyl nitrosoethylurea.

            J.Cell Biol. 1974. 61: 318-413; J.Biol.Chem. 1977. 252: 2143-2153.

Morphology:  fibroblast-like

Mode of cultivation: monolayer

Conditions for cultivation: medium - DMEM

serum -   FBS 20%

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

EDTA  0.02% (1:3), split ratio 1:3 - 1:10, optimal population density 1.0-5.0104 cells/cm2

cryoconservation - growth medium, 8% DMSO, 1.0x106 cells/ml in ampule

Viability after cryoconservation:     95% (0 passage, dye trypan blue)

Sterility: tests for bacteria, fungi and mycoplasma were negative

Species: karyological analysis

Karyology:   2n= 40, variability in the range between 60-76 chromosomes,

modal number of chromosomes 64-67, number of polyploid cells 8%.

Other properties:

synthesis of adenylate and creatine phosphokinases, acetylcholine receptors.

Possess many properties characteristic of smooth muscle.

Applications:  acetylcholine receptors study.

Collections: ATCC CRL 1443; ECACC 86093001; SPBIC.