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Chemical and Biological Properties of B16 Murine Melanoma Cells Grown in Defined Medium Containing Bovine Serum Albumin¹

Department of Biological Chemistry and the Specialized Cancer Research Center, The Milton S. Hershey Medical Center, The Pennsylvania State University, Hershey, Pennsylvania 17033

The addition of 1% (w/v) bovine serum albumin (BSA) to the F12 medium utilized for the growth of the B16 melanoma cells significantly stimulated the growth of this cell line. The synthesis of mucopolysaccharides and sialoglycopeptides in this medium is identical with that in Eagle's minimal essential medium with Earle's balanced salt solution supplemented with 2 mM L-glutamine, twice the recommended concentration of vitamins, nonessential amino acids, sodium pyruvate, and 10% (v/v) fetal calf serum. Cell volume and morphology did not change significantly under the different growth conditions and tumorigenicity, as assayed by injection of cultured cells into syngeneic animals, was not decreased. Analysis of the BSA used indicated the presence of a sialoglycoprotein contaminant. This sialoglycoprotein contaminant was present in all lots examined and contains N-acetyl- and N-glycolylneuraminic acid, mannose, galactose, and glucosamine. The sialoglycoprotein can be removed by chromatography on acetate form anion-exchange resin at pH 4.3. F12 media containing the purified BSA plus selenite and the sodium salts of palmitic, oleic, and linoleic acids supported growth of the melanoma cells to the same extent as did the media containing unpurified BSA, indicating that the sialoglycoprotein has no role in sustaining the growth of the cells.

¹ Supported in part by USPHS Grant CA15483 and Grant 1 P30 CA18450-01, awarded by the National Cancer Institute, Department of Health, Education, and Welfare.

² To whom requests for reprints should be addressed.

Received 2/10/77. Accepted 8/30/77.