L-M (TK-, APRT-)

Origin: mouse, connective, derived from NCTC clone 929.

            Submitted Institute of Biochemistry, Martinsried, FGR.

Morphology:  fibroblast-like

Mode of cultivation: monolayer

Conditions for cultivation: medium - DMEM

serum -  FBS 10%

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

EDTA  0.02% (1:3), split ratio 1:5 - 1:10, optimal population density 2.0-3.0104 cells/cm2

cryoconservation - growth medium, 10% FBS, 5-10% DMSO, 1.8106 cells/ml in ampule

Viability after cryoconservation:     68% (0 passage, dye trypan blue)

Sterility: tests for bacteria, fungi and mycoplasma were negative

Species: karyological, isoenzymological (LDH, G6PD)  and immunological analysis

Karyology:   2n= 40, variability in the range between 46-51 chromosomes,

modal number of chromosomes 49, number of markers - 9 metacentrics (routine dye).

Plating efficiency:    25%

Tumorigenicity:  non tumorigenic

Other properties:

deficient in thymidine kinase and adenine phosphoribosyl

transferase (resistant to 5-bromodeoxyuridine and 8-azaadenine.

Retrovirus type A production

Applications: virology, somatic cell genetics, cell biology.

Collections:   SPBIC.