Selection of a chemically defined medium for culturing adult newt forelimb regenerates

Three commercially available tissue-culture media were evaluated for their ability to support continued growth and differentiation of 14-day regenerates of adult newt forelimbs. Serums, embryo extracts, egg ultrafiltrates, and antimicrobial agents were avoided in this analysis. The hormones insulin and L-thyroxine were added to these chemically defined media to enhance continued cellular metabolism and growth. The optimum conditions appeared to be cultivated at 25 degrees C (pH 7.2 to 7.4) in media osmotically adjusted to conditions approximating amphibian blood values (i.e. 225 m0SM for 199, 244 m0SM for CMRL-1066, and 262 m0SM FOR L-15).