Integrated cell culture bioengineering

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Mammalian cell culture has enormous utility and potential for the production of recombinant proteins, vaccines, viral vectors and cells as such, like stem cells and tissues. Culture processes for these cells are very complex since their culture media is often made of more than 40 costly components, and their cell internal reaction network is composed of thousands of signalling and metabolic steps, often cell-type and time dependent. Many primary cultures, such as haematopoietic, muscle and skin cells, still need foetal bovine serum (FBS), an undefined mixture of biological source, as well as a layer of feeder cells to grow normally. Although many supplements have been proposed to replace FBS and feeder layers, they often don't allow the production of cells that are functional for therapy. Our lab specializes in integrating a set of approaches to study and improve culture conditions for mammalian cells. 1) Statistical design of experiments (DOE) is applied to evaluate the individual and synergistic effects of panels of potential factors. 2) Sequential FBS fractionation is developed which, combined to DOE, allows to discover entirely new active factors and combination thereof. 3) FBS fractions obtained in (2) are analysed by mass spectroscopy to identify molecules supporting this activity. 4) Automated microscopy and image analysis rapidly evaluate cell response to the tested factor, allowing the evaluation of cell growth, and most interestingly, cell shape through morphometric parameters which are clear indicators of the cell state. 5) Fluid handling station is being designed to address the specific needs of our research. A last approach, 6) Genome-scale metabolic modelling using the latest technologies to build mathematical models. Genomic databases and estimation of each network flux rate by large-scale linear optimization will be combined with our experimental platform to form a complete high-throughput evaluation system for mammalian cell culture processes. This research program proposes pursuing the development, integration and improvement of this innovative experimental/theoretical platform and applying it to the culture of keratinocytes used for the treatment of severe burn, diabetic foot ulcers and oral mucosa defects.