The establishment of cell lines plays a important role for the study on cellular and molecular biology of cancer. lt is well known that serum free defined media is useful for the establishment of cell lines. The effect of twelve factors used as a constituents of serum free defined media, ACL-4, on seven colon cancer cell lines were tested using MTT [3-(4-5-dimethythiazol-2-yl)-2, 5-diphenyl tetrazolium bromide] assay. Our results suggest that among twelve factors tested, transferrin and insulin and bovine serum albumin were very important factors for the growth of colon cancer cell lines. The concentration of transferrin (10 ug/ml) and insulin (20 ug/ml) in ACL-4 media was optimal to culture of the twa colan cancer cell lines, but one line, NCI-H716, graws better in higher concentration of insulin (100 ug/ml).
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