Selected nutrients reduce the pyruvate requirement for survival in vitro of chick central nervous system neurons

Central nervous system neurons, cultured as monolayers at low density, need exogenous pyruvate for their survival. The pyruvate concentrations required by embryonic day 8 (E8) chick forebrain neurons are reduced substantially by other low molecular weight agents present in Dulbecco's modified Eagle's medium (DMEM) and in astroglia‐conditioned Eagle's basal medium (EBM). To examine the nature of these pyruvate‐sparing molecules, E8 chick forebrain neurons were cultured in EBM containing a concentration of pyruvate too low to support neuronal survival, unless sparing agents were supplied. This stragegy permitted the identification of the pyruvate‐sparing DMEM constituents, omission of which had led to neuronal loss. Pyruvate sparing required a mixture of (i) serine, (ii) pyrioxal (or riboflavin and biotin), and (iii) six essential amino acids: arginine, cystine, leucine, isoleucine, tyrosine, and tryptophan (each replaceable by corresponding amino acids, except tryptophan). All pyruvate‐sparing contributors were optimally needed at or near their DMEM concentrations.