CHEMICALLY DEFINED MEDIA FOR CULTIVATION OF LONG-TERM CELL STRAINS FROM FOUR MAMMALIAN SPECIES.

Abstract A number of new cell lines have been grown in chemically defined medium NCTC 109 free of any exogenous protein. The list in this laboratory now includes fibroblast-like and epithelial-like cells, normal and malignant, and in origin from the mouse, hamster, monkey and human. The lines growing in medium NCTC 109 were then tested for their growth in simpler media of the NCTC series: 117, 126, 131, 132 and 133. Medium 117 lacks the six coenzymes and deoxyguanosine, deoxyadenosine, 5-methyl-cytosine, and sodium acetate present in medium 109. Media 126, 131, 132 and 133 lack all of these except sodium acetate; nine vitamins, glutathione, cysteine and sodium glucuronate were also omitted. Medium 131 is identical with medium 126 except that the former contains vitamin B 12 . Media 132 and 133 are identical with medium 126 except that 132 contains inositol and 133 contains both inositol and vitamin B 12 . Two human lines grew readily in media 117 and 126, as did also the monkey kidney cells and some of the mouse cell strains. All mouse cells tested grew in medium 117 except the mouse liver cells. Hela and mouse liver cells showed anomalous growth behavior in media 117, 126, 131 and 132; these two strains are still under test. When mouse cells did not growin media 126 and 131 it was usually because they required inositol. Of all of the cell strains conclusively tested only one strain of mouse cells and the one Chinese hamster strain required vitamin B 12 for continuous growth at least when thymidine and deoxycytidine were included. This report, timely because of the increased significance and more widespread use of various chemically defined media, also describes improvements in quality control and simplified preparation methods for the NCTC series of media. Our experience strongly emphasizes the necessity for high quality in the chemicals and for the meticulous preparation and storage of such media when used without any protein supplements.