Aplicació d'estratègies d'enginyeria metabòlica en la millora de processos basats en el cultiu in vitro de cèl.lules animals

This proposal presents an approach to the improvement of procesess based on animal cells culture, based on the results and technology developed in previous projects. The starting point is based on the fact that even for improved procesess by means of control systems and perfusion opoeration, important bottle-necks are observed, being the most important that of cell death in the cultures. This phenomena is associated to the deregulated metabolism and to the programmed cell death process, that can be triggered by multiple causes. The project approaches this limitations combining simultaneously macroscopical aspects of the culture and genetic engineering aspects of the cells (metbolic engineering). Specefically, it is proposed to develope a bi-phasic culture strategy, in wich a first growth phase is decoupled from a second cyclostatic phase, maintaining the product formation, and with long duration. The project proposes different alternatives to ahieve such transition, both for fed-bacht and perfusion cultures. In any case, these strategies must be complemented with a genetic modification of the cells, specifically introducing a limitations to the apoptosis progress, minimizing cell death along the second phase of the culture. Two representative cell lines of the industrial operation are used: hybridomas and CHO, producing respectively a monoclonal antibody and recombinant t-PA.