Periodic Reporting for period 2 - ENLIGHT (ENable LIGHT- and synthetic biology-driven volumetric bioprinting of functional human tissues)
Okres sprawozdawczy: 2022-05-01 do 2023-10-31
To achieve this goal, ENLIGHT will:
- Develop a novel, highly efficient route to generate multiple subsets of endocrine pancreatic cells from stem cells/induced pluripotent stem cells
- Develop materials able to substitute the native pancreas extracellular matrix, to allow to nurture the engineered stem cells in 3D
- Develop a novel, ultra-fast volumetric bioprinting technique to sculpt these cells and materials into large-sized pancreatic organoids
- Enable the long term culture of such 3D organoids in a perfusion system.
- Investigate the potential of these organoids as drug testing platform and develop a strategy for their use a transplantable cell therapy.
In parallel, several gelatin-based hydrogels formulations have been developed, and a base design that allows the formation of 3D clusters of engineered beta-like cells, to preserve the identity and function of iPSC-derived pancreatic islets as well as the formation of interconnected capillary networks from a co-culture of endothelial HUVEC cells and mesenchymal stromal cells has been defined. The material can be shaped via volumetric printing, and their functionality via the embedding of ECM matrix components. To sculpt these cell-laden materials, a new volumetric, tomographic 3D printing technology has been developed, enabling the rapid fabrication of centimeter scale constructs in less than 30 seconds. Several strategies have been developed to ensure the encapsulation of high cell densities contextually to a high shape fidelity and printing resolution. Light-based printing can be impaired by opaque media and scattering caused by the cells in the printable hydrogel., Data-driven scattering correction by frequency boosting allows to fabricate unobstructed vasculature models in preliminary experiments at 4 million cells/mL. In addition, refractive-index matching with biocompatible contrast agents improves print resolution in preliminary experiments in presence of 10 million cells/mL. Biomaterial design and software can be used to compensate for scattering and achieve high resolution in cell-laden hydrogels. . We also demonstrate multiple strategies to produce geometrically complex multi-material prints, which comprise also spatial patterning of multiple cell types.
A fully functioning set-up for the sterile perfusion of geometrically complex, centimetre scale constructs printed from hydrogels displaying low mechanical properties was designed and tested. Perfusion culture of endothelial cells seeded in bioprinted channels was achieved. Bioprinting of iPSC derived islet organoids (obtained by chemical differentiation) is possible, and to date, culture up to 21 days, with nearly 100% viability has been performed.
Moreover, the dissemination framework for ENLIGHT was established in the first six months of the project. This included the website, logo, and social media platforms, along with a data management plan. The ENLIGHT partners were active in dissemination and communication efforts, including press releases by international media outlets, the development of project videos, presentations at virtual, global scientific conferences, and the publishing of peer-reviewed publications. In the first 6 months, the project management structure and consortium guidelines were established and distributed to the consortium via the PM Handbook. The project management strategy has thus far been sufficient to maintain lines of communication between partners and to ensure that any potential issues are addressed early and proactively.