Bioengineering for Cell-based Therapies

Bridging bioprocess engineering and stem cell biology to accelerate the development of advanced cell therapies.

Bioengineering for Cell-based Therapies at iBET

At iBET, we employ our over 30 years of expertise to improve the manufacturing of cell therapy products for clinical applications.

Our interdisciplinary teams combine upstream and downstream bioprocess development technologies with state-of-the-art bioanalytical tools to support our partners at any stage of their cell therapy product development pipeline. Our approach privileges the intersection of process intensification, scalability, cost-effectiveness, and quality control to ensure potency of the final product.

Process design for cells as therapeutic products

In collaboration with academic and industrial partners, we focus on bioprocess development and intensification for scalable production of (stem-derived and immune) cell therapy products with improved potency.

We design scalable bioprocesses for autologous and allogeneic cells (human cells, including pluripotent stem cells, mesenchymal stem/stromal cells, CAR-T cells) and their derivatives (differentiated cells and extracellular vesicles, EVs), both at upstream and downstream levels.

We employ our expertise in single-cell and 3D culture strategies, bioreactor technology and rational development of downstream purification methods. Our efforts are directed towards the use of cGMP-compatible manufacturing platforms, based on disposable technology (bioreactors, perfusion systems and membrane chromatography tools) and on the use of xeno-free components (e.g. culture medium, matrices).

Expansion and differentiation of hPSC-derivatives for engineered tissues/organs

Ongoing R&D projects tackle the improvement of hPSC expansion and differentiation/maturation into functional cell types for the development of bioengineered tissues/organs for heart and liver regenerative medicine.

In collaboration with academic and industrial partners, we are developing biology-inspired protocols, combining the knowledge of cell/developmental biology with the expertise of bioprocess engineering, to generate hPSC-derived cardiac, liver and kidney cell types.

Key steps include the modulation of critical environmental factors, namely the metabolic substrate, dissolved oxygen, co-culture with other hiPSC-derivatives, biomaterials, and extracellular matrix (ECM) components.

Characterization and (cryo)preservation of cell therapy products

Our bioprocess development is integrated with robust bioanalytical tools, which support the monitoring of cell therapy products’ quality attributes. We employ “omics”-based bioanalytical tools and process analytical technologies (PAT), including Dielectric and Raman Spectroscopy, for improved process understanding and optimization. More traditional analytical assays are also applied whenever applicable.

We further develop (cryo)preservation protocols for human (stem) cells (adult, embryonic and induced pluripotent stem cells) to facilitate their storage and transportation. Strategies for the short- term (i.e. cold preservation) and long-term (i.e. cryopreservation and vitrification) of cells as single cells, 2D monolayers or 3D cultures are also available, using, for example, cell entrapment in a permeable hydrogel matrix.

Related Technologies

Cell Culture Approaches

We possess expertise in a diverse range of cell culture approaches, including monocultures and co-cultivation strategies; cell immobilization in microcarriers/scaffolds; cell aggregates; Tissue explants; Microencapsulation (cell immobilization in inert biocompatible matrices, such as alginate, or defined ECM components).

Bioreactor Platforms

We employ bioreactor technology for bioprocess development and intensification. Available systems include: glass (80 mL to 5 L) and stainless-steel stirred tank bioreactors (5 to 50L) and disposable systems (15ml to 50 L). With these technologies) we can finely control and monitor environmental parameters (pO2, pH and perfusion rate) to specifically optimize cell expansion and differentiation steps with maximal yields.

Multi-Omics Tools

We employ multi-“omics” approaches (proteomics, transcriptomics, metabolomics and fluxomics) for a detailed molecular-level characterisation of cellular function, focused on aspects including cell survival, metabolic function, cell proliferation and/or differentiation/maturation.

Metabolic Characterization

We employ a wide range of equipment to assess metabolic function: CEDEX Bio for detection of metabolites, HPLC, NMR, LC-MS and GC-MS analytical techniques, integrated through metabolic flux analysis methodologies.

Imaging

To assess cell morphology, functionality, and marker expression we employ state-of-the art imaging techniques. Confocal microscopy (2-photon, spinning disk, light sheet) is used for live fluorescence-based assays and immunofluorescence characterization. Scanning and transmission electron microscopy are used for morphological analysis.

Downstream Processing

We design integrated purification approaches combining membrane-based technologies (e.g. expanded bed chromatography) and filtration-based strategies (dead-end filtration and tangential flow filtration using hollow fibers and flat sheet cassettes) for downstream processing of cells and EV's. Key steps of the process include harvesting, concentration/volume reduction, cell washing and removal of undesired cell populations and other impurities.

Highlights

Development of a novel CAR-T therapy against pediatric leukemia

We joined a European consortium, led by OneChain Immunotherapeutics (OCI), and funded by the European Innovation Council (EIC) Transition Challenge (Grant Agreement nº 101113067), working to develop a promising CAR T-cell based therapy against T-cell acute lymphoblastic leukaemia.

The novel therapy will use a subtype of T-cells (gamma delta), which are allogenic, eliminating donor dependency, and prone to scalable manufacture, therefore representing a promising off-the-shelf cell product. The carxall project (www.carxall.com) started in April 2023 and, in the next 3 years, the iBET team will be playing a key role in translating the production process to the industrial level.

EIC Transition Challenge

“Large-scale” production of hiPSC-derived cardiac cells in bioreactors

Within “BRAV∃” project and in collaboration with University of Navarra (UNAV, Spain) and University Medical Center Utrecht (UMCU, Netherlands), scalable bioprocesses for generation of cardiac cells derivatives (cardiomyocytes, cardiac fibroblasts, cardiac endothelial cells) from hPSC are being developed.

Within “BRAV∃” project and in collaboration with University of Navarra (UNAV, Spain) and University Medical Center Utrecht (UMCU, Netherlands), scalable bioprocesses for generation of cardiac cells (cardiomyocytes, cardiac fibroblasts, cardiac endothelial cells) from hPSC are being developed.

These cells are produced in stirred-tank bioreactors (up to 2L scale) and after comprehensive characterization of quality attributes, will be used to seed the BioVAD (Biological Ventricular Assisted Device) and will define the new functional cardiac tissue to be tested in large animal models. Know more: BravE (projectbrave.eu)

EU Horizon 2020

Enhanced bioprocess control to advance the manufacture of Extracellular Vesicles

In collaboration with University of Navarra (UNAV, Spain), we developed a scalable manufacturing workflow to maximize the production of extracellular vesicles (EV) from mesenchymal stem/stromal cells in stirred-tank bioreactors.

The integrated bioprocess increases EV production 5-fold compared to conventional methods, with optimal isolation using tangential flow filtration and size exclusion chromatography. We also explored the potential of Raman spectroscopy to continuously track metabolite levels in stirred-tank bioreactors, contributing to streamline the selection of optimal EV collection timepoints. Importantly, MSC-derived EV maintained their quality attributes and stimulated angiogenesis in vitro, therefore highlighting the therapeutic potential of EVs manufactured following a scalable and tightly controlled bioprocess. The work, developed under the scope of the CARDIOPATCH project, funded by Interreg-Sudoe, was selected as the cover feature for the special issue: “Advanced Cell Technologies” in Biotechnology and Bioengineering (Marta H Costa et al., 2023).

Interreg-Sudoe

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Related Teams

Stem and Immune Cells Bioengineering Lab

Margarida Serra

Lab Head, Stem and Immune Cells Bioengineering Lab | Advisor to the Executive Committee

Bridging engineering and stem cell biology to accelerate the development of advanced cell therapies.

Translational Immunology Lab

Nádia Duarte

Senior Scientist, Translational Immunology Lab

Strengthening immunology expertise at iBET, with a focus on developing tools and strategies to test and potentiate the effectiveness of advanced cell immunotherapies