Periodic Reporting for period 1 - BLUES (Bioprocesses for metabolite production from marine invertebrate cell lines)
Reporting period: 2024-01-01 to 2025-06-30
The BLUES project aims to expand the production of valuable bioactive compounds from marine invertebrates by developing new cultivation systems. The focus is on four phyla: Porifera, Cnidaria, Echinodermata, and Chordata. This initiative offers a sustainable alternative to wild harvesting by optimizing production yields. After years of research, BLUES partners have achieved a significant breakthrough: the creation of the first continuous marine sponge cell line. This discovery demonstrates that sponge cells from various species can divide rapidly.
The ultimate goal is to design a pathway for industrial bioprocessing using these invertebrate cell lines as a foundation for producing unique, high-value marine compounds. These novel bioprocesses not only resolve the bottleneck of stock production but also contribute to a more sustainable, circular economy. The technology developed within the project will enable the production of valuable marine natural products through bioprocesses, thereby contributing to the blue bioeconomy.
The ultimate goal is to design a pathway for industrial bioprocessing using these invertebrate cell lines as a foundation for producing unique, high-value marine compounds. These novel bioprocesses not only resolve the bottleneck of stock production but also contribute to a more sustainable, circular economy. The technology developed within the project will enable the production of valuable marine natural products through bioprocesses, thereby contributing to the blue bioeconomy.
The BLUES project has been running for 18 months and has produced a series of deliverable defined in the grant proposal. These include, but are not limited to:
• Assessing the proliferation rates of 10 sponge species, which produce valuable metabolites, to determine which 2 species to select for the development of immortal cell lines
• Developing a novel photo-bioreactor for the culture of corals with a day and night cycle
• Dissection methodologies for the sea cucumber C. frondosa to isolate and extract cells from specific layers of the body wall
• Optimization of extraction and culture protocols for various sponge, coral, sea cucumber cells and tunicate cells
• Fabrication of 3D scaffolds, three in total, made from marine derived collagen for scale-up of marine invertebrate cultures
• Scale-up of G. cydonium and G. barretti cultures from T25 flasks (~10mL) to 2 x 125mL custom made bioreactors using 3D scaffolds made of marine derived collagen
• Quantification of Frondoside A yields from frozen and dried C. frondosa. Analysis was conducted on both the whole organism and different layers of the body wall. This informed the layer of interest for cell culture
• Quantification of Stevensine yields from frozen and dried A. corrugata
• Improvement of methods for sponge hybridoma development using both poly-ethylene glycol and electrofusion
• Assessing the proliferation rates of 10 sponge species, which produce valuable metabolites, to determine which 2 species to select for the development of immortal cell lines
• Developing a novel photo-bioreactor for the culture of corals with a day and night cycle
• Dissection methodologies for the sea cucumber C. frondosa to isolate and extract cells from specific layers of the body wall
• Optimization of extraction and culture protocols for various sponge, coral, sea cucumber cells and tunicate cells
• Fabrication of 3D scaffolds, three in total, made from marine derived collagen for scale-up of marine invertebrate cultures
• Scale-up of G. cydonium and G. barretti cultures from T25 flasks (~10mL) to 2 x 125mL custom made bioreactors using 3D scaffolds made of marine derived collagen
• Quantification of Frondoside A yields from frozen and dried C. frondosa. Analysis was conducted on both the whole organism and different layers of the body wall. This informed the layer of interest for cell culture
• Quantification of Stevensine yields from frozen and dried A. corrugata
• Improvement of methods for sponge hybridoma development using both poly-ethylene glycol and electrofusion
BLUES will have a significant impact through publications that will be generated during the project, presentation of findings at internationally recognized colloquia, interactions with related projects such as COMBO (101135438), and licensing novel technologies developed. We have already characterised the biomass from C. frondosa and A. corrugata, identified and conducted initial testing of deep-eutectic solvents (DES) for extraction of secondary metabolites from tissue samples. The consortium is also developing and testing novel DES combinations for multi-product biorefinery processes to further improve biomass, and eventually cell biomass, usage – thereby improving the value obtained from the process.
We have also tested the use of scaffolds - made of natural marine-based components such as collagen from fish skin – for the ability to support the attachment and growth of marine sponge cells G. cydonium and G. barretti in a bioreactor of 125mL. This is the first and largest bioreactor cultivation of marine-sponge cells in history, making this a groundbreaking achievement. Given that this achievement has come so early in the project, we aim to build upon this success and further improve the cell culture via media optimisation, scaffold optimisation and increasing and improving the bioreactor scale and functionality.
The first two needs to ensure uptake - further research and demonstration - are the main ones being targeted. There is potential to leverage IPR support, which is being investigated further by the consortium leader, Wageningen University, as the project develops and produces patentable outcomes. It is currently too early in the project to predict what additional measure will be necessary, but this will be monitored throughout the course of the project.
We have also tested the use of scaffolds - made of natural marine-based components such as collagen from fish skin – for the ability to support the attachment and growth of marine sponge cells G. cydonium and G. barretti in a bioreactor of 125mL. This is the first and largest bioreactor cultivation of marine-sponge cells in history, making this a groundbreaking achievement. Given that this achievement has come so early in the project, we aim to build upon this success and further improve the cell culture via media optimisation, scaffold optimisation and increasing and improving the bioreactor scale and functionality.
The first two needs to ensure uptake - further research and demonstration - are the main ones being targeted. There is potential to leverage IPR support, which is being investigated further by the consortium leader, Wageningen University, as the project develops and produces patentable outcomes. It is currently too early in the project to predict what additional measure will be necessary, but this will be monitored throughout the course of the project.