Optimization of growth performance of freshly induced carrot suspensions concerning PMP production

Carrot suspension cultures are efficient production systems for plant-made pharmaceuticals (PMP); however, frequently, the reduction of transgene expression in long-running transgenic cell lines limits the productivity. Freshly induced carrot suspensions have been identified for a constant and high biomass production with optimization of culture conditions and cultivars. A small-volume monitoring method was adapted to freshly induced carrot suspension that minimized the heterogeneity of freshly induced meristematic carrot cell lines. Using this system combined with the adaptation of growth conditions, we identified the cultivar Rote Riesen as having the highest increase in dry biomass with an average of 6.5 g/L weekly, stable and consistent after 8 wk. As a model PMP, VP60-the capsid protein from the rabbit hemorrhagic disease virus-was produced in a carrot suspension with a maximal accumulation of 2.4 mu g/g dry mass. The developed method guarantees comparable investigations of different transgene expression in various freshly induced small-volume carrot cell lines