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The cells are 5-bromo-2'-deoxyuridine resistant (BUDR, BrdUrd) and thymidine kinase (TK) deficient, HAT sensitive.

Tested and found negative for ectromelia virus (mousepox).

1. Remove and discard culture medium.
2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
3. Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
   Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
5. Add appropriate aliquots of the cell suspension to new culture vessels.
6. Incubate cultures at 37°C.

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Chen TR. Cytogenetics of somatic cell hybrids. II. Modal karyotypes and mosaicism in mouse mutant cell lines: LM(tk-), A9, and Rag. Cancer Genet. Cytogenet. 24: 95-107, 1987. PubMed: 3466673

Hsu TC, Somers CE. Properties of L cells resistant to 5-bromodeoxyuridine. Exp. Cell Res. 26: 404-410, 1962. PubMed: 14449548

Boone C, et al. Assignment of three human genes to chromosomes (LDH-A to 11, TK to 17, and IDH to 20) and evidence for translocation between human and mouse chromosomes in somatic cell hybrids (thymidine kinase-lactate dehydrogenase A-isocitrate dehydrogenase-C-11, E-17, and F-20 chromosomes). Proc. Natl. Acad. Sci. USA 69: 510-514, 1972. PubMed: 4110482

Peterson JL, McBride OW. Cotransfer of linked eukaryotic genes and efficient transfer of hypoxanthine phosphoribosyltransferase by DNA-mediated gene transfer. Proc. Natl. Acad. Sci. USA 77: 1583-1587, 1980. PubMed: 6929511

Higazi AA, et al. Soluble human urokinase receptor is composed of two active units. J. Biol. Chem. 272: 5348-5353, 1997. PubMed: 9030610

Carballo E, et al. Bone marrow transplantation reproduces the tristetraprolin-deficiency syndrome in recombination activating gene-2 (-/-) mice. J. Clin. Invest. 100: 986-995, 1997. PubMed: 9276715