Production of human epidermal growth factor (hEGF) by in vitro cultures of Nicotiana tabacum: effect of tissue differentiation and sodium nitroprusside addition

Abstract

The aim of this work was to study the production of human epidermal growth factor (hEGF) by in vitro cultures of Nicotiana tabacum. Suspension and hairy root cultures expressing hEGF were established and the time course of growth and hEGF production were evaluated. In hairy root cultures, hEGF content was about 2 µg g -1FW, with a productivity of 15.9 µg l-1d-1 whereas in cell suspensions, a specific production of 0.48-0.53 µg g -1FW and a productivity of 31 µg l-1 d-1 were obtained. From these results, it is clear that hEGF synthesis and accumulation depend on tissue differentiation. In order to increase hEGF production, cell suspension cultures were treated with casaminoacids and sodium nitroprusside (SNP, a nitric oxide donor). SNP treatment resulted in an increase of hEGF productivity of 48% compared with control cultures. In conclusion, cell suspension and hairy root cultures established from transgenic N. tabacum were able to produce hEGF and the concentration and yield obtained depended on the level of tissue differentiation and the treatment applied.