LoopOfFun is a platform for the rapid development of industry-scale, one-step, simple casting-based manufacturing processes

for materials based on renewable and locally sourced raw materials.

Designing and Manufacturing of stools with open- and closed-loop controlled Structure. Photos: Johannes Falkenstein

The first printing attempts using GeSiM mbH Nanoplotter 2.1 Bioink CMC solution with mycelium A. Niger. Photos IGB

About

Fungi comprise approximately 100 000 described species to date. The real total is estimated to be in the millions. They are amazing factories, producing numerous bioactive metabolites of therapeutic interest. The EU-funded LoopOfFun project has recognised their potential in yet another innovative area – as part of engineered living materials (ELMs), with control of mechanical and structural properties. The project will identify fungi gifted with superior abilities for materials synthesis and harness them for synthetic biology-based programming. The programming will be accomplished via a novel automatic robotised platform that will develop the fungi into ELMs, based on iterative design-build-test-learn cycles. The outcomes will then support rational design of such materials.

PROJECT Description

In an interdisciplinary approach, LoopOfFun will develop a conceptual and technological framework for the efficient and effective development of fungal-based living materials with controlled mechanical and structural properties. This framework is based on four pillars: 

  • The development of the synthetic biological tools for writing mechanical and structural material properties

  • The identification of filamentous fungi with naturally evolved superior properties for material synthesis as well as accessing them for synthetic biology-based programming 

  • The development of the 4D Explorer, an automated, robotized platform for the efficient and effective development of engineered living materials based on iterative design-build-test-learn cycles 

  • A holistic approach for designing the final products based on the novel opportunities of LoopOfFun advances.  

 Following this framework, LoopOfFun will produce materials with defined mechanical and structural properties. The intrinsic features make the materials robust towards environmental fluctuations while aligning material properties with usage characteristics.  

The development and applicability of the novel materials will be shown by producing two different demonstrator materials at cm-m scale, one as structural material and the other one for pollutant degradation. The framework developed here as well as its underlying routines can generically be applied for the design and optimization of ELMs made of other organisms and matrices. 

Penicillium rubens with GFP-SKL targeted to the peroxisomes. Van der Lende et al, 2002, Biochim. Biophys. Acta

The LoopOfFun Consortium has internationally leading expertise and access to state-of-the-art technology in all fields relevant to the EIC-2021-Pathfinder Challenges-01-05.

—> INM – Leibniz-Institut für Neue Materialien, Saarbrücken, Germany

—> Kemijski Institut, Slovenia

—> Rijksuniversiteit Groningen, Netherlands

—> Fraunhofer Gesellschaft zur Forderung der Angewandten Forschung EV, Germany

—> ASA LUMA/Arles, France

Consortium

are experts in Molecular Optogenetics as well as in the synthesis of biohybrid materials with informationprocessing properties based on synthetic biological sensors and switches. In addition, the research team at INM has gained significant expertise in the design and application of AcAl-based interkingdom communication systems. On the technological side, INM has state-of-the-art equipment for engineering, producing, purifying, modifying and characterizing proteins such as fermenters and automated chromatography systems. Further, W. Weber is member of the Freiburg Materials Research Centre providing access to state-of-the-art hardware for materials characterization (rheometer, universal testing machines, EM etc.). This leading expertise in both, synthetic biology and biohybrid materials excellently position INM for the development of the defined circuit for porosity as well as for leading the development of the HLM-based filter material with closed-loop controlled anti-clogging features. Visit the group here.

The formal coordinator was Albert-Ludwigs-Universität Freiburg from November 01, 2022 to June 30, 2023 due to the transition of the principal investigator and his research team to the INM - Leibniz-Institut für Neue Materialien.

INM – Leibniz-Institut für Neue Materialien, Saarbrücken, Germany (the coordinator)

Dr. Trong Hoang Phan, Postdoc Researcher

Prof. Dr. Wilfried Weber, Scientific Director,

Principal Investigator

Prof. Dr. Roman Jerala, Head of Department,

Principal Investigator

Prof. Dr. Arnold Driessen, Principal Investigator

is an interdisciplinary creative campus located in Arles (F) and gathering thinkers, artists, researchers, and scientists to question the relationships between art, culture, environment, education, and research. One of Luma-Arles’ programs, Atelier Luma is a research design platform that researches the ecosystems of the Camargue bioregion and develops applications such as fungi-grown materials while promoting the local natural and cultural resources. Atelier Luma is committed to balancing social and economic development. Its multi-disciplinary team explores possible scenarios for transforming consumer cultures and existing systems of production. As a locally focused experimental lab, Atelier Luma also participates in knowledge sharing networks and in developing pilot projects around the world. Visit here or back to Consortium

SAS LUMA/ARLES, France (LUMA Arles)

Dr. Ha T. M. Pham, Project Coordinator

KEMIJSKI INSTITUT, Slovenia

are experts in molecular microbiology of fungi, and in particular with respect to the development and application of synthetic biology tools including Crispr/Cas9 genome editing and the use of the MoClo language to construct genetic parts. These technologies are used to optimize fungal hosts strains for the production of secondary metabolites, remove undesired genes, as well as the integration of heterologous small molecule biosynthetic pathways for biotechnological applications. Visit the group or back to Consortium

Carlotta Borgato, Project Manager / Designer

are experts in the design of protein-based switches and the design of 3D protein structures using coiled-coil motifs. They further invented informationprocessing circuits and modules in mammalian cells based on proteases and coiled-coils and optimized systems for efficient secretion of proteins. They have further access to state-of-the-art hardware for the analysis of protein and materials structures including cryo-electron microscopy as well as diverse biophysical techniques such as DLS, MALS, SAXS, AFM, SPR and ITC. Further, technologies for single molecule microscopy as well as FRAP and FCCS are available. These leading skills especially on designing coiled-coil-based molecular tools combined with the technological equipment, position NIC as the preferred partner in developing the molecular closed-loop control circuits for stiffness adaptation in response to mechanical stress. Visit the group here or back to Consortium

Dr. Weijun Zhou, Post-doc Researcher

RIJKSUNIVERSITEIT GRONINGEN, Netherlands

is the leading organization for applied research in Europe, bridging industrial and academic R&D environments. The group led by Achim Weber provides leading end-to-end solutions, from the laboratory to pilot-scale in the 3D printing of biological and bio-based materials. They have leading expertise in materials science, interfacial engineering and material characterization. Especially, they possess established pipelines for the development of bio-inks for 3D printing of different organisms, materials and matrices as well as of state of-the-art equipment for characterizing the produced materials with regard to mechanical and structural properties. This expertise positions A. Weber perfectly for leading the development of the 4D Explorer platform based on the modification of 3D printing hardware. Further, Kai Sohn is an expert in environmental and functional genomics, (meta)genomics/transcriptomics including the construction and maintenance of custom-based databases and respective search tools and strategies. His sequencing and annotation pipeline will be an important asset for the genomic characterization of the material-assembling fungal strains. See more here or back to Consortium

Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V., Germany

Dr. Joachim Weber, Deputy Head of the Innovation Field, Principal Investigator

Jan Boelen, Artistic Director Principal Investigator

Dr. Kai Sohn, Head of innovation field in-vitro diagnostics

Klemen Mezgec, PhD Researcher

LoopOfFun is part of a portfolio of projects funded under the Engineered Living Materials Pathfinder Challenge by the European Innovation Council and started on November 2022.

With this Pathfinder ELMs Challenge the EIC seeks to seize the opportunity to position strategically Europe at the forefront of the ELMs field. This Pathfinder Challenge aims to overcome the technological challenges to harness the engineering potential of nature for materials’ production. The specific objectives of Pathfinder ELMs Challenge are to support the development of new technologies and platforms enabling the controlled production of made-on-demand living materials with multiple predictable dynamic functionalities, shapes, and scales; and to build a community of researchers and innovators in ELMs.

ELMs projects funded from the EIC Open calls actively contribute to the Portfolio activities with the aim to advance the scientific and technological development of ELMs and promote its dissemination across Europe, increase the visibility of the ELMs community internationally by sharing knowledge and building partnerships, engage with regulatory bodies to address ELMs portfolio needs, address ethical, legal and social aspects through early engagement with policymakers and the public, and to assess and address the need for standardization in the ELMs portfolio, identify barriers to the adoption and commercialization of ELMs and engage with stakeholders, guided by responsible research and innovation methods.”

—> BioRobot-MiniHeart

—> Fungateria

—> Furoid 

—> LoopOfFun

—> NextSkins

—> PRISM-LT

—> SUMO

LIST OF PROJECTS

Engineering a swimming bio-robot and a living human mini-heart.

Coordinator: University of Twente (NL)

Project partners: 4

Key-words: tissue engineering, biosensing, stem cells, cardiovascular diseases, physiology

Project description: Manufacturing our very own hearts is just a heartbeat away, literally. Engineers are joining forces with biologists to make biological heart robots. The EU-funded BioRobot-MiniHeart project is developing a vascularised beating mini-heart. In parallel, the team is designing a self-propulsion swimming bio-robot created by assembling human cardiac cells into 3D tissue structures; using sacrificial moulding and high-resolution 3D bioprinting.

The mini-heart and the bio-robot will provide scientists with a more realistic human cardiac model in vitro and an appropriate tool to assess cardiotoxicants’ presence in the environment. We expect this innovation to help speed up the development of heart disease cures. Visit Website or Back to the List of Projects

BioRobot-MiniHeart

Combining fungi and bacteria into novel biomaterials

Coordinator: Royal Danish Academy – Architecture, Design, Conservation (DK)

Project partners: 6

Key-words: bacteriology, synthetic biology, mycology

Project description: Engineered living materials (ELMs) are composed of living cells endowed with unique properties and functions. ELMs have received significant attention in materials sciences due to their tuneability and potential for sustainable production. Funded by the European Innovation Council, the Fungateria project aims to generate an innovative portfolio of ELMs that combine fungi with bacteria.

Growing the vegetative part of the mushroom—the mycelium—on different organic substrates is the most common way of producing fungi-based materials. The project will combine the mycelium with bacteria that serve as a chassis for sensor-containing genetic circuits. The resultant ELMs will exhibit advanced functionalities and inducible degradation when no longer needed. Visit Website or Back to the List of Projects

Fungateria

Up-scaled, continuous production of artificial hair, fur, and wool follicles.

Coordinator: BIOFABICS LDA (PT)

Project partners: 3

Key words: textiles, ecosystems, additive manufacturing, transplantation

Project description: The mission of FUROID is to enable the animal-free production of hair (humans), fur (endangered animals), and sheep wool. Our innovative approach will use a combination of nanostructured scaffolds (RESPILON), hair/wool/fur organoids (GENEUS), and automated biofabrication technologies (BIOFABICS) to develop engineered living fur (ELF). We plan to design continuous and scalable technology that can dominate the market by 2030.

Our ambition to develop hybrid living materials-based products will deliver a set of distinctive properties:

·       Animal free-production of fur (ELF) and wool (ELW) without environmental, biodiversity and ethical concerns

·       Gene-encoded traceability system endangered species unnoticed, uncontrolled and unpunished endangered species poaching

·       Novel vacuum-assisted 3D printing technology overcoming the speed of SoA 3D bioprinters

·       Continuous manufacturing using an automated production system

·       DBTL platform for rational design and product optimisation

·       Living human hair follicles (ELH) for transplantation as a medical and psychological treatment for alopecia

·       Fur and wool textiles for the apparel industry in rolls without size limitation

·       Improved properties of engineered fur/wool textiles due to nanofiber membranes with regulated water permeability

The project will strengthen the portfolio of Engineered Living Materials, developing scalable and generalisable technology for forming textured materials in the roll-to-roll process from mammalian cells. Visit Website or Back to the List of Projects 

Furoid

LoopOfFun

Fungi-based engineered living materials with controllable properties.

Coordinator: Albert-Ludwigs-Universitaet Freiburg (ALU-FR) (DE) until 30.06.2023. From 01.07.2023 Leibniz-Institut fuer neue Materialien gemeinnutzige GmbH (INM) (DE) is the coordinator because of the switch of the PI and his team from ALU-FR to INM

Project partners: 5

Key-words: Mycology, electrical engineering, sensors

Project Description: Fungi comprise approximately 100 000 described species to date. The real total is estimated to be in the millions. They are amazing factories, producing numerous bioactive metabolites of therapeutic interest. The EU-funded LoopOfFun project has recognised their potential in yet another innovative area – as part of engineered living materials (ELMs), with open- and closed-loop control of mechanical and structural properties. The project will identify fungi gifted with superior abilities for materials synthesis and harness them for synthetic biology-based programming. The programming will be accomplished via a novel automatic robotised platform to develop the fungi into ELMs based on iterative design-build-test-learn cycles. The outcomes will then support the rational design of such materials. Visit Website or Back to the List of Projects

Living therapeutic and regenerative materials with specialised advanced layers.

Coordinator: Delft University of Technology (NL)

Project partners: 3

Key-words: bacteriology, dermatology, biomolecules, ceramics

Project Description: Compared to conventional materials, biomaterials in living organisms possess specific architecture and organisation: and often exhibit multiple functions. Εngineered living materials (ELMs) have emerged at the junction of synthetic biology and material science to produce materials with improved functionality because of the living organisms within them.

Funded by the European Innovation Council, the NextSkins project is inspired by the structure and function of the many layers of skin. Researchers will mimic the specialised skin arrangement to make two engineered living materials: one with a therapeutic role to treat skin diseases and one with a purpose to be used as a protective garment in sports. Visit Website or Back to the List of Projects

NextSkins

Living tissue manufacturing using symbiotic materials

Coordinator: IN society (IT)

Key-words: bacteriology, stem cells, bioprinting,

Project Partners: 6

Project Description: The EU-funded PRISM-LT project will use a hybrid living materials concept to create a flexible platform for living tissue manufacturing. The innovative bio-ink will contain stem cells integrated into a supporting matrix with engineered helper bacteria or yeast cells. The bioprinting process will produce a 3D patterned structure where stem cells could be induced to differentiate into different lineages. The directed stimulation of differentiating stem cells will force them to produce lineage-specific metabolites for sensing by the designer helper cells. The helper cells within the platform will then enhance localised lineage commitment to sustain differentiation stability. The project aims to implement this strategy for the development of two symbiotic materials designed for biomedical and food applications, respectively. Visit Website or Back to the List of Projects 

PRISM-LT

Supervised morphogenesis in gastruloids as an alternative to conventional single-tissue organoids.

Coordinator: Oslo University Hospital (NO)

Project partners: 7

Key words: artificial intelligence, developmental biology, stem cells, physiology

Project Description: The lack of realistic in vitro organ models that faithfully represent in vivo physiological processes is a major obstacle affecting the biological and medical sciences. The current gold standard is animal experimentation, but it is increasingly evident that these models mostly fail to recapitulate human physiology. Moreover, animal experiments are controversial, and it is a common goal in the scientific community to minimise the use of animals to a strictly necessary minimum.

The emergence of stem cell-engineered organ models called organoids represents the only viable alternative to animal research. However, current organoid technology is yet to produce the larger physiologically relevant organ models that the medical sciences need. Specifically, current organoids are too small, not vascularised and lack the 3-dimensional organisation found in vivo.

In this interdisciplinary project, we aim to challenge all these limitations using the recently developed gastruloid technology guided by cutting-edge bioengineering and artificial intelligence.

Gastruloids are formed by initiating the very early developmental processes and develop along a highly coordinated three-axial process that closely resembles mammalian embryogenesis. They can establish several organ precursors simultaneously, thus constituting relevant improvements over conventional single-tissue organoids.

To harvest the potential of gastruloid technology, we will first implement extensive sequencing and imaging experiments to optimise the developmental trajectory of gastruloids for organ inductions. We will then build these datasets into a multimodal data matrix to identify gastruloid candidates for cardiovascular and foregut development. Candidates with substantial vasculogenesis will be chosen for later vascularisation by anastomose with endothelial cells.” Visit Website or Back to the List of Projects

SUMO

  • Kick Off

    2 November 2022

  • The 1st annual meeting

    25 - 27 June, 2023

    Atelier Luma, Arles, France

The 1st annual meeting

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The LoopOfFun consortium met for the first time in person at Atelier LUMA in Arles, France. It was a fantastic meeting with lots of shared science on, interesting perspectives, and helpful exchanges at a lovely and attractive Gehry tower.

Dissemination

Under Construction