Bioprocess design for the manufacture of cells and extracellular vesicles with therapeutic potential
Currently the production of cells and derived extracellular vesicles (EVs) is performed in laborious and time-consuming static culture systems using serum-containing media and with no control of culture conditions. The main focus is the development of efficient, scalable and cost-effective production processes for the large-scale ex vivo expansion of human adult cells (namely hematopoietic stem/progenitor cells, mesenchymal stromal cells, NK cells, skeletal and smooth muscle cells and endothelial cells) and the production of their EVs in controlled bioreactors. Moreover, the identification of the critical quality attributes (CQAs) of cells/EVs and the establishment of a standardized methodology to measure those is being developed. Although a variety of techniques have been used to characterize cells/EVs, this area is still far from accomplishing the needed standardization, especially for EVs.
Development of downstream processes for the isolation and purification of cells and extracellular vesicles
Cells and EVs have been typically purified at a laboratory scale using methods based on their properties/characteristics (e.g. size, morphology, density, composition or immunological features) such as velocity and density gradients; agglomeration or precipitation; and adsorption chromatography (commercially available kits). Scalable downstream processes for the isolation and purification of cells/EVs comprising unit operations that are robust, selective and cost-effective are being developed and integrated with the upstream processing.
Integration of culture monitoring platforms for bioprocess control and optimization
Smart monitoring platforms are being developed to ensure continuous culture assessments mandatory to develop robust bioprocesses essential to implement novel cell therapies. This includes the implementation of process analytical technologies (PAT) along upstream and downstream processes, in order to identified critical process parameters and critical material attributes and to study their impact in the critical quality attributes (CQA) of cells/EVs.
Team:
PhD holders
Ana Fernandes-Platzgummer, Smart bioprocess design for the manufacture of cells and extracellular vesicles with therapeutic potential (Keywords: Stem Cells, Extracellular vesicles Bioprocess design, Bioreactors, Process control, Scalable production)
Raquel Cunha, Development of a scalable manufacturing process for therapeutic human mesenchymal stem/stromal cell-derived exosomes (Keywords: mesenchymal stromal cell, conditioned medium, extracellular vesicles, scalable manufacturing, therapeutic potential)
Sara Morini, Biomanufacturing platforms for the large-scale expansion of non-parenchymal cells for liver bioengineering purposes (Keywords: Non-parenchymal cells, Bioreactors, Liver bioengineering)
PhD students
André Branco, Design and Operation of a Prototype Bioreactor for the Expansion and In Situ Selection of Human Hematopoietic Stem/Progenitor Cells (Keywords: Hematopoietic Stem/Progenitor Cells, Bioreactor Design, Process Optimization)
Master students
Catalina Cepeleaga, Breaking down mesenchymal stromal cells: uncovering the potential of decellularized extracellular matrix (Keywords: Mesenchymal stromal cells, extracellular matrix, decellularization)
Diogo Estrela, Delineation of optimal culture conditions for therapeutic human mesenchymal stromal cell-derived extracellular vesicles production in dynamic cultures (Keywords: Mesenchymal stromal cells, Extracellular vesicles, Dynamic culture)
Key collaborators
Pedro Baptista (Aragon Health Sciences Institute, Zaragoza), Kamilla Swiech (University of São Paulo), Cecília Calado (ISEL/IPL), Carla Lilaia (HSFX/CHLO), Ana Azevedo (BERG-IBB/IST), Pedro Fonte (BERG-iBB/IST), Stefan Spichiger/Ferdi Caglayan (C-CITSwitzerland), William Milligan (AventaCell Biomedical Corp. Ltd) and Ricardo Egea (Bionet Servicios Técnicos, S.L).