Effect of zinc on insulin-free hybridoma culture

Abstract

The murine hybridoma CRL1606 was found to be dependent on insulin for proliferation in serum-free conditions. A novel insulin-free medium, developed by zinc-supplementation, could support cell growth and antibody production to the same extent as the original insulin-supplemented medium. No adaptation into the new medium was necessary. The effects of these media on the transcriptional response of the cells were investigated using a 7524-element microarray based on the Compugen basic mouse oligolibrary. Analysis via pathway visualization suggested that zinc could have insulin-mimetic effects in insulin-free, zinc-supplemented CRL1606 culture via the up-regulation of PUR-1 and CD19 (which then imparts survival signals via the Pi3k-dependent pathway), as well as through the down-regulation of the apoptotic adaptor FADD (that eventually inhibits apoptosis via the caspase 8 cascade). Taken together, the microarray results suggest that Pi3k may be important in integrating zinc-induced signaling with the insulin-mimetic effects observed in zinc-supplemented insulin-free CRL1606 cultures.