Periodic Reporting for period 1 - ENLIGHT (ENable LIGHT- and synthetic biology-driven volumetric bioprinting of functional human tissues)
Período documentado: 2021-05-01 hasta 2022-04-30
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 investigated, and a base design that allows the formation of 3D clusters of engineered beta-like cells, 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 107 cells/mL. Biomaterial design and software can be used to compensate for scattering and achieve high resolution in cell-laden hydrogels.
A first functioning set-up for the sterile perfusion of geometrically complex, centimetre scale constructs printed from hydrogels displaying low mechanical properties was designed and tested. The prototype will be refined and used with the pancreatic constructs in the coming reporting periods. At the same time we started to develop methods to readily observe the differentiation of human pluripotent stem cells into pancreatic progenitors and then to alpha/beta cells via fluorescent reporters., to facilitate cell differentiation and health readout analysis within the bioprinted organoids.
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.