LONG-TERM CULTIVATION OF A HUMAN COLONY-STIMULATING FACTOR-PRODUCING CELL-LINE IN A PROTEIN-FREE CHEMICALLY DEFINED MEDIUM

  • 1 January 1984
    • journal article
    • research article
    • Vol. 44  (10) , 4503-4506
Abstract
To examine whether a human colony-stimulating factor (CSF)-producing cell line, T3M-1, can propagate and secrete CSF without protein supplements, long-term cultivation of the cells was carried out in a protein-free chemically defined medium. By the use of stepwise decreases in the fetal bovine serum concentration, continuous growth of T3M-1 cells was established in a protein-free F-10 medium. The cells designated T3M-1-T2 were propagated in this medium for 5 yr. The population-doubling time of the cells was .apprx. 30 h. Addition of serum stimulated the cell growth (population-doubling time, 17 h) but did not increase the saturation density. The cells secreted large amounts of CSF (2000 colonies stimulated by 1 ml of the conditioned medium). Addition of serum to the culture increased CSF activity in the conditioned medium (3400 colonies/ml). A human cancer cell line, T3M-1-T2, could be propagated in a protein-free chemically defined medium and secrete large amounts of CSF. The cells will serve as an excellent model for better understanding of the cell growth and production of CSF in the absence of any serum contamination.