AN IMPROVED CHEMICALLY DEFINED BASAL MEDIUM (CMRL-1415) FOR NEWLY EXPLANTED MOUSE EMBRYO CELLS

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Abstract
An improved chemically defined medium, CMRL-1415, has been devised by testing the response of trypsinized, newly explanted mouse embryo cells in stationary cultures to various modifications of an earlier medium, CMRL-1066. The improvements are attributed to changes in amino-acid levels, in the vitamin-coenzyme composition, and to an enhanced buffering capacity resulting from the use of free base amino-acids, galactose and pyruvate, together with greatly reduced levels of glucose and sodium bicarbonate and the inclusion of both monobasic and dibasic sodium phosphate in the ratio 1:4. When the medium is equilibrated with 5% CO2 In air, an initial pH of 7.2[long dash] 7.4 is achieved, with excellent buffering capacity. CMRL-1415 contains 50 ingredients (9 fewer than CMRL-1066) and is prepared from six stable stock concentrates. By omitting sodium bicarbonate (to give CMRL-1415-ATM), the medium may be used in un?ealed cultures in free gas exchange with air. CMRL-1415 and CMRL-1415-ATM are intended for use with and without serum protein and other supplements; and by preparing them double strength they may be combined with agar or other gelling agents to provide a semi-solid substrate.