Project

• Less than 10% of global transplantation needs are met: 1 in 4 patients in the waiting list die or become too ill to undergo transplant surgery. In the EU, 1,000 patients die every year, waiting for a donor liver.
• Whole organ bioprinting hampered by low output of cell/spheroid production, lack of appropriate bioinks, low viability and vascularisation post-printing.

Bioprinting presents a promising approach for creating organs from scratch, yet, it faces significant hurdles due to technical and biological challenges, combined with lacking standardized procedures and materials. NEOLIVER forms the largest knowledge base on bioprinted liver constructs as it builds on a previous European project ORGANTRANS, which successfully used organoids and supporting cells to generate thousands of multicellular spheroids, mimicking in vivo tissue complexity. The spheroids served as the basis for extrusion-based bioprinting, resulting in small bioprinted constructs successfully validated in immune-deficient mice. We have identified specific barriers in whole-organ bioprinting that NEOLIVER will tackle before reaching the patient, while promoting acceptance by end-users, regulatory bodies, and the general public. To advance whole-organ tissue engineering, we need to produce dense bioprinted tissue with a vascular bed that can be scaled up and integrated into a standardized and automated process. Barriers will be systematically addressed by integrating advanced expertise in cell biology, biomaterials, vascularization, bioprinting, automation, and clinical translation. We will generate new bioprinting tools (LIFT) to create dense tissue, and core-shell extrusion based bioprinting for vessel formation. By using spheroids, combined with novel synthetic hydrogels, we can study the interconnectivity of different cell types and create fully vascularised bioprinted constructs. As a pre-clinical model, size-appropriate and fully functional bioprinted liver constructs will be transplanted in immune-deficient pigs, with a subsequent development of a clinical validation plan for first-in-human trials. NEOLIVER will develop Good Manufacturing Practice (GMP)-conform protocols and processes, encompassing cell sourcing, mass-scale expansion, bioprinting of liver constructs using synthetic hydrogels, vascularization, and tissue maturation for transplantation purposes

1. Establish and expand cell sources for tissue engineering.
2. Develop encapsulated multicellular spheroids and synthetic hydrogel precursors.
3. Create a laser bioprinting platform for high-speed printing of large dense bioprinted tissue.
4. Develop novel vascularization processes for transplantable bioprinted constructs.
5. Develop novel vascularization processes for transplantable bioprinted constructs.
6. Demonstrate the functionality of the liver construct in vivo in a large animal model.
7. Ensure the clinical and regulatory readiness and acceptability of the product.

Despite advances in organ transplantation technology, there is still a huge shortage of transplantable organs. Yearly, 25% of patients with end-stage liver disease on the donor waiting list die, emphasizing the need for alternatives to organ donations, such as bioprinting. Bioprinting presents a promising approach for creating organs from scratch, yet, it faces significant hurdles due to technical and biological challenges, combined with lacking standardized procedures and materials. In NEOLIVER, we will develop large, dense, and vascularized fully functional bioprinted constructs suitable for transplantation. We will achieve this by establishing a GMP-conform manufacturing line for standardized production, ensuring unparalleled quality and safety for future patients. More specifically, by using patient-derived organoids and supporting cells including endothelial cells, we will generate millions of multicellular spheroids as building blocks for bioprinting. Through laser induced forward transfer (LIFT) bioprinting techniques we will create a vascularized liver construct via precise spatial deposition of spheroids and vessels at high density. By integrating this technology with extrusion-based bioprinted vessels for blood supply, we will generate the world’s first autologous bioprinted liver, ready for transplantation. To show the safety and efficacy, we will transplant the bioprinted liver constructs in immune-deficient pigs. This, combined with a clinical validation plan, upscaling strategy and Health Technology Assessment (including patient acceptance), will prepare the bioprinted liver constructs for first-in-human trials. Thus, NEOLIVER presents a disruptive alternative to donor organs for patients dealing with end-stage liver disease.

• Societal: Transplant waiting time decreased from 15 months to 11.5 weeks, avoiding waiting list mortality of 1,000 patients/year in the EU in the long-term. Reduction of adverse effects caused b y long-term immunosuppression. Up to 3,700 patients/year in the EU gain additional 11.5 quality adjusted life years.
• Scientific: Large scale bioprinting units for liver constructs expanded to other solid organs, e.g. pancreas, heart, kidney.
• Economic: EU becomes a global leader in regenerative medicine tools for tissue engineering and transplantation; Increased economic efficiency thanks to patients return to the labour market and reduced post-transplantation costs.
• Technological: Novel technologies for upscaling, spheroid production (including encapsulation), bioprinting and maturation of bioprinted tissue constructs under standardised conditions that can progress the technologies to an industrial scale.