Adaptation, Growth, and Chromosomal Analysis of HEp-2 Cells in Chemically Defined Medium2

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Abstract
The tissue culture cell line HEp-2 has been adapted to growth in several chemically defined NCTC media. Continuous growth of these cells in medium NCTC 135 as static and fluid suspension cultures was excellent. Partial nutritional analysis on the subline of HEp-2 adapted to growth in chemically deAned medium NCTC 135 showed the cells fo have a vitamin 812 and an i-inositol requirement for successful growth. HEp-2 cells growing in the chemically deAned medium NCTC 135 and in NCTC 135 supplemented with 10 percent v/v fetal calf serum were successfully frozen, stored, and recovered From liquid-nitrogen temperatures. No significant alterations in the cells recovered from liquid-nitrogen temperature were noted in the parameters studied. Chromosome analysis before, during, and after the adaptation of HEp-2 cells to chemically defined medium NCTC 135 revealed: 1) a stemline shift From 77 when grown in medium 199 with 10 percent calf serum to 70 when grown in chemically defined medium NCTC 135, 2) the retention of the 3 old marker chromosomes and the appearance of new marker chromosomes in the cells during the adaptation to growth in chemically defined medium NCTC 135, and 3) a narrow range of chromosome numbers in cells grown in beth medium NCTC 135 with 10 percent v/v fetal calf serum and medium NCTC 135 with no serum supplement.