Facci L, Skaper S D, Varon S
J Neurosci Res. 1985;14(3):293-302. doi: 10.1002/jnr.490140302.
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 strategy 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) pyridoxal (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.