PRInted Symbiotic Materials as a dynamic platform for Living Tissues production

Project Information

PRISM-LT

Grant agreement ID: 101070913

DOI

10.3030/101070913

EC signature date 16 June 2022

Start date 1 November 2022

End date 31 October 2027

Funded under

• The European Innovation Council (EIC)

Total cost

€ 2 805 403,82

EU contribution

€ 2 805 403,82

Coordinated by

IN SRL
Italy

Periodic Reporting for period 2 - PRISM-LT (PRInted Symbiotic Materials as a dynamic platform for Living Tissues production)

Reporting period: 2023-11-01 to 2025-02-28

Summary of the context and overall objectives of the project

Recent breakthroughs in cell engineering and additive manufacturing have opened new frontiers for developing Engineered Living Materials (ELMs), constructed partially or entirely from living cells. These ELMs, characterized by a hierarchical arrangement of different cell lineages, exhibit multifunctional properties that often surpass those of both natural living materials and traditional non-living materials. However, realizing the full potential of living tissue manufacturing requires overcoming significant challenges, such as achieving controlled printability on a commercially viable scale and speed.

In response to these challenges, the EU-funded PRISM-LT project emerges as a pioneering initiative. Its primary objective is to establish a versatile platform for the 3D bioprinting of living tissues, endowed with dynamic functionalities and predictable shapes. Drawing inspiration from the mechanisms governing natural tissue development, PRISM-LT sets out to engineer heterogeneous 3D printable living materials capable of self-assembly into intricate living tissues. This ambitious endeavor spans scales ranging from sub-millimeters to centimeters.

PRISM-LT will assess the efficacy of its platform through the utilization of two distinct symbiotic materials, each directed towards specific applications: one in the biomedical realm (utilizing organoids as in-vitro models for biomedical research) and the other in the domain of food-related innovations (cultivating marbled cultured meat).

Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far

In the latest phase, the project made major strides in developing engineered helper cells to support tissue formation for medical and food applications.

i) Tissue Engineering
Bacterial helper cells (E. coli) were engineered to produce and secrete important growth factors to support bone formation.
Two innovative secretion strategies were established: one that anchors the protein to the cell membrane, and another that enables secretion into the surrounding medium.
To prevent overgrowth and ensure safe co-culture with stem cells, a new type of non-dividing helper cell called a minicell was developed. These minicells remain active and capable of protein production without multiplying, making them ideal partners in tissue culture systems.

ii)Cultured Food
Engineered yeast cells were developed to secrete muscle- and fat-promoting growth factors.
A sensing system based on lactic acid detection was introduced, allowing yeast to respond to signals from stem cells and adjust their activity accordingly.
A second approach is being explored to make yeasts responsive to physical cues like the stiffness of the surrounding material, helping further tailor the support they provide during early tissue development.

iii)Bioprinting and Bioinks
Major advances were made in bioink formulation and bioprinting techniques, as two strategies were explored:
1) Direct mixing of stem cells with soft or stiff bioinks to guide tissue formation.
2) Encapsulation of cells in microcapsules of defined stiffness, which are then printed into 3D structures.
A range of new tunable bioinks were developed using materials, optimized for both mechanical properties and biological function.
New bioprinting protocols and software tools were introduced, enabling precise patterning of soft and hard regions, mimicking natural tissue structures like bone and fat.
Finally, an innovative mechanical testing method (based on Brillouin microscopy) was developed to verify the printed structures' properties.

Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far)

The PRISM-LT project not only addresses the technological hurdles of controlled printability but also aligns with broader societal and scientific needs. By creating a nexus between cutting-edge advancements in biofabrication and tissue engineering, PRISM-LT aims to contribute significantly to the understanding and application of Engineered Living Materials. The envisioned impacts of the project extend beyond the laboratory, holding the promise of transformative applications in various fields, including biofabrication, tissue engineering, and beyond.

While tackling specific biomedical and food applications the platform is very flexible and has several applications. During the project PRISM-Lt will put its platform to the test by developing biomaterials for i) organoids as in vitro models for preclinical research, which could aid in discovering and testing of new drugs or therapies, and ii) synthetic meat that closely mimics its natural counterpart, incorporating typical marbling, texture, nutritional values, and safety.

Identifying these impactful results, the potential for further uptake and success becomes apparent. Areas requiring attention may include additional research to fine-tune and expand the platform's capabilities, demonstrations to showcase its real-world applicability, access to markets and financial support for scaling up production, commercialization efforts to bring these innovations to market, intellectual property rights (IPR) support to protect the project's innovations, internationalization strategies for global reach, and a supportive regulatory and standardization framework to facilitate widespread adoption. These considerations collectively form a roadmap for ensuring the sustained impact, scalability, and success of the PRISM-LT project.

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