THE GROWTH RESPONSE OF MAMMALIAN CELLS IN TISSUE CULTURE TO l-GLUTAMINE AND l-GLUTAMIC ACID

Abstract

Twelve n-amino acids (arginine, cystine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, tyrosine, and valine) have been found essential for the growth of two mammalian cell lines in tissue culture, a mouse fibroblast' (strain L) (1) and a human uterine carcinoma cell (strain HeLa) (2).The minimal vitamin requirement of these cell lines has also been defined (3).As will be shown here, under the conditions of the present experiments, glutamine proved similarly essential for the survival and growth of both cell lines.In a medium containing the twelve amino acids previously shown to be essential, the seven demonstrably essential vitamins, glucose, electrolytes, and serum protein, both the mouse fibroblast and the human carcinoma cell degenerated and died unless the medium was supplemented with glutamine.The quantitative aspects of this glutamine requirement, and the limited degree to which it could be satisfied by glutamic acid, are described. Methods and MaterialsThe methods of maintaining stock cultures of the two cell lines, of setting up replicate cultures with the various experimental media, and of cell enumeration to evaluate the growth response have been described in previous papers dealing with the amino acid and vitamin requirements of the mouse fibroblast (1, 3) and the HeLa carcinoma cell (2, 3).A helpful modification in the cell-counting, procedure has been the use of disodium Versenate (disodium salt of ethylenediaminetetraacetic acid) to disperse the cells, instead of mechanically scraping them from the surface of the culture flask.The flask was drained, and 3 ml. of fresh medium at pH