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Subcultures are prepared by scraping. Remove one-half of the old medium, add fresh, dislodge cells from the floor of the flask, aspirate and dispense into new flasks.

Complete growth medium described above supplemented with 5% (v/v) DMSO.  Cell culture tested DMSO is available as ATCC Catalog No. 4-X.

Mizel SB, et al. Characterization of lymphocyte-activating factor (LAF) produced by the macrophage cell line P388D1. I. Enhancement of LAF production by activated T lymphocytes. J. Immunol. 120: 1497-1503, 1978. PubMed: 149171

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Evans VJ, et al. Chemically defined media for cultivation of long-term cell strains from four mammalian species. Exp. Cell Res. 36: 439-474, 1964. PubMed: 14242231

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Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

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Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.