A-9

Origin: mouse C3H/An, subcutaneous adipose connective tissue, derived from NCTC 929.

            Proc.Natl.Acad.Sci. 1963. 50: 568; Nature 1964. 202: 1142; Am.J.Human Gen. 1974. 26: 273.

Morphology:  fibroblast-like

Mode of cultivation: monolayer

Conditions for cultivation: medium - EMEM or DMEM

serum -   FBS 10%

other components -   NEAA 1% (EMEM)

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

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

cryoconservation - growth medium, 10% 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 and isoenzymological (LDH, G6PD) analysis

Karyology:   2n= 40, variability in the range between 52-57 chromosomes,

modal number of chromosomes 54-55, number of markers - 1 (routine dye),

number of polyploid cells 1.8%.

Other properties:

deficient in hypoxanthine phosphoribosyltransferase, resistant to 8 - azaguanine and 6 - thioguanine

May be heterozygous for the ability to synthesize active inosinic acid phosphorylase.

Applications:  metabolism, genetics of somatic cells.

Collections: ATCC CRL 6319; ECACC 84011426; SPBIC.