Introduction

The oceans cover over 70% of Earth’s surface and 95% of Earth’s biosphere and contain a wealth of biological diversity with more than 200,000 described species of marine invertebrates and algae and millions of microbial species (United Nations 2018). This vast biological diversity is accompanied by an equally remarkable chemical diversity of secondary metabolites. One evolutionary explanation for this vast array of chemical compounds is that many marine invertebrates are sessile as adults and have evolved in habitats which are often more challenging, compared to their terrestrial counterparts. This has resulted in the evolution of novel metabolic pathways and the production of structurally diverse chemicals that have a wide variety of ecological roles (Paul et al. 2011, Puglisi et al. 2014, Puglisi et al. 2019, Carroll et al. 2020), including compounds to reduce predation, enhance larval settlement or reproduction, contribute to direct inter- and intraspecific competition, reduce palatability or nutrient uptake by predators, protect against parasites, viruses and pathogens, as well as regulate host microbiomes.

Such compounds are being exploited for potential industrial uses, such as pharmaceuticals, cosmetics, nutritional supplements, molecular probes, enzymes, anti-foulants, fine chemicals and agrichemicals (Rotter et al. 2021, Rotter et al. 2024). For example, the first marine (sponge)-derived products to become clinically available were the result of a modest research programme in the 1950s (Bergmann and Feeney 2002), which led to the synthesis of the nucleosides vidarabine (Ara-A), an antiviral drug and cytarabine (Ara-C), an anti-cancer drug. Further efforts slowly developed over the next two decades and, in the 1980s, major initiatives in marine drug discovery were initiated. Several marine-derived molecules since then have become clinically available as pharmaceuticals (Rodrigues et al. 2019, Saeed et al. 2021). Specifically, from 1990–2019, more than 15,000 new natural products from marine invertebrates have beenreported (Calado et al. 2022, Brück et al. 2023), but only a small percentage of these compounds has been studied for potential uses (Li et al. 2018), in part, because of the supply bottleneck. The most prolific sources are Porifera (47.2%) and Cnidaria (35.3%), with Echinodermata and Chordata contributing 7.7% and 6.1%, respectively (Jeon et al. 2015, Kobayashi and Ishibashi 2000, Carroll et al. 2022).

Marine organisms produce a wide range of bioactive metabolites with potential human health applications, including those that have cytotoxic, antioxidant and antimicrobial properties. Marine invertebrates share a surprisingly high level of homology with more developed metazoans (Gamulin et al. 1994, Schäcke et al. 1994). Genes encoding for toll-like receptors (Cetkovic et al. 2004, Rodrigues et al. 2019), src-type proteins (Cetkovic et al. 2004, Ma et al. 2014), Ras-like small GTPases (Cetkovic et al. 2007), Bcl-2 homology domains (Wiens et al. 2000), death domains (Müller et al. 2000, Wiens et al. 2000), tyrosine kinases (Muller et al. 1999), mortalin (an HSP70 member that induces senescence in immortalised mouse fibroblasts) (Ben-Hamo et al. 2018) and survivin (a regulator of proliferation and apoptosis) (Luthringer et al. 2010), have all been identified in lower invertebrate taxa, like sponges (phylum Porifera) and show significant homology to mammalian genes. Antimicrobial peptides (AMPs) have gained attention as natural compounds possessing biological activities and significant potential for medical applications. AMPs are crucial effectors of the innate immune system of marine organisms and regulate proliferative activities and protective processes. In marine invertebrates, humoral factors encompass AMPs and peptides dissolved in coelomic fluid or haemolymph (Guryanova et al. 2023). The need for more research, characterising antimicrobial compounds from echinoderms, for example, as potential sources of novel pharmacologically relevant compounds, has already identified. (Hanifaturahmah et al. 2024, Zhao et al. 2024, Atanassova et al. 2024).

Marine-derived bioactive compounds can also address the rising demand for alternative nutraceuticals (Eroldoğan et al. 2022), pharmaceuticals and cosmeceuticals (Rotter et al. 2024), as well as for new bio-based products (Rotter et al. 2021). Additionally, marine biomass can be utilised to create bulk commodities like biofuels and bioplastics (Tennakoon et al. 2023). The sustainable use of marine resources and development of biomolecules and polymers are key aspects of marine biotechnology.

Despite the vast potential for marine compounds to act as new pharmaceuticals, cosmetics, nutritional supplements, molecular probes, catalysts, anti-foulants, fine chemicals, agrichemicals and other innovative bioproducts, the major bottleneck to the development of this biotechnological sector remains the limited supply of raw materials. Harvesting wild organisms from the marine environment is not ecologically sustainable, as many of the novel marine compounds are found only in minute quantities and are often only synthesised under special conditions (Brück et al. 2023). Furthermore, a wide range of bioactive compounds/chemicals cannot be reproduced through chemical synthesis, aquaculture or wild collections. The testing of a single drug may require several kilograms of a specific bioactive metabolite for preclinical and clinical stages, potentially demanding thousands of tonnes of fresh material (Calado et al. 2022). Clearly, such biomass quantities cannot be harvested from nature without risking the continued existence of the respective species, resulting in many promising marine-derived natural compounds that can no longer be pursued for product development.

The BLUES consortium has been assembled to tackle this research challenge regarding the development of cell cultures from marine invertebrates. It is comprised of experts in cell culture of the four taxa of invertebrates (Porifera, Cnidaria, Echinodermata, Urochordata); marine invertebrate-microbe symbiosis; product down-stream processing; process intensification; and communication, exploitation and dissemination (Fig. 1). The consortium consists of nine partners from eight countries (Table 1) and is funded by a Horizon Europe Circular Economy and Bioeconomy Sectors (HORIZON-CL6-2023-CIRCBIO-01) grant, project number 101134820. The 4-year project was initiated on 1 January 2024 and is coordinated by Wageningen University (WU).

Table 1.

Download as

CSV

XLSX

BLUES consortium members.

Partner	Country
Wageningen University	Netherlands
Moreforsking AS	Norway
Israel Oceanographic and Limnological Research Limited	Israel
Matis OHF	Iceland
Universidade Do Minho	Portugal
Universita Degli Studi Di Genova	Italy
Geonardo Kornyezetvedelmi Terinformatikai Es Regionalis Projektfejleszto Korlatolt Felelossegu Tarsasag	Hungary
Cellex SRL	Italy
University of New South Wales	Australia

Figure 1.  

Graphical representation of the BLUES Work Package structure and interdependencies.

Scientific Challenges and BLUES Innovation for the Blue Bioeconomy

Novel Extraction Methodologies for Secondary Metabolites

Secondary metabolites extracted from wild-harvested marine invertebrates are typically present in low concentrations. These low yields make extraction unsustainable, as the biomass required would place significant pressure on natural populations, threatening species’ viability. This creates a harmful cycle in which increasing demand leads to overharvesting, further endangering marine biodiversity. Moreover, variability in metabolite content due to environmental factors makes wild harvesting an unreliable and inconsistent source for large-scale production.

Deep eutectic solvents (DESs) have emerged as promising green solvents, gaining considerable attention in both academia and industry in recent years due to their low toxicity, non-volatility, low vapour pressure, high boiling point and ease of preparation. DES are typically formed through mixtures of Bronsted or Lewis acids and bases, usually involving at least one hydrogen bond donor (HBD) and a hydrogen bond acceptor (HBA), resulting in a eutectic system (Fig. 2). By varying the type and ratio of HBD and HBA components, DES can be tailored to exhibit specific physicochemical properties, making them highly versatile for targeted applications. The particularly notable subclass of natural deep eutectic solvents (NaDES) (Duan et al. 2016) are composed of naturally occurring primary metabolites that play an important role in cellular processes (e.g. sugars, carboxylic acids, amino acids and alcohols). These components are generally abundant and non-toxic, rendering NaDES both environmentally sustainable and safe for use in food and pharmaceutical extractions. Recently, DESs have been explored as alternative solvents for extraction of bioactive compounds, offering a “greener” substitute for conventional organic solvents. They have shown potential in the extraction of a variety of metabolites (e.g. flavonoids, proteins, lipids and polysaccharides) from sources like plants, microalgae and lignocelluloses (Duan et al. 2016). Due to their highly tunable nature, DESs also open the door to integrated, multi-product biorefinery process, thereby increasing the sustainability of the extraction processes.

Figure 2.  

Schematic representation of a eutectic point on a two-component phase diagram (Smith et al. 2014).

Recently, we developed a complete extraction process using DES for isolating lipids from the microalga Nannochloropsis oceanica (Lo et al. 2021), alginate and pigments from Brown seaweed (Hiemstra et al. 2025, Hiemstra et al. 2026, Reynaga-Navarro et al. 2026). Within WP5, the focus is on advancing downstream processing methods; an often overlooked yet critical aspect of marine biotechnology. Given the typically low concentrations of target metabolites in marine biomass, conventional methods often have a significant environmental footprint and generate substantial waste. To address this, WP5 aims to develop mild, efficient and environmentally sustainable extraction protocols that not only reduce the ecological impact, but also enable a multi-product approach. By thoroughly exploring the composition of each cell line, WP5 seeks to recover multiple valuable compounds without compromising their quality, ensuring that co-products remain intact and suitable for high-value applications. This integrated strategy maximises the use of marine biomass, improves overall resource efficiency and enhances the economic viability of marine bioprocessing.

Increasing Product Yields

As noted earlier, extraction yields of marine-derived metabolites are often low. The BLUES project addresses this challenge through two key strategies: increasing product yields and intensifying the overall production process. To enhance metabolite yields, several approaches will be implemented, including the optimisation of culture media tailored to species-specific needs, investigating the genetic mechanisms underlying metabolite biosynthesis, the use of hybridoma technology and exploring co-culture systems involving invertebrate hosts and their algal or microbial symbionts.

One major strategy for culturing sponge cells has involved adapting existing mammalian cell culture media by supplementing them with salts to mimic sea water (formulations such as Marine M199) and incorporating additional components, such as growth factors to stimulate cell proliferation (Pomponi 1997, Pomponi et al. 1998a, Pomponi et al. 1998b, Willoughby and Pomponi 2000, Krasko et al. 2001, Zhang et al. 2002). Recently, we optimised amino acid concentrations, based on metabolic activities for the sponge Dysidea etheria, leading to the development of a new medium, called ‘M1’ (Munroe et al. 2019). Following this, we observed rapid cell division in nine additional sponge species (Conkling et al. 2019). In particular, we established finite cell lines for three species of the same genus, Geodia cf. gibberosa, G. neptuni and G. barretti, which could be subcultured up to five times and achieved a maximum of seven population doublings (Conkling et al. 2019). Building on this, we formulated an enhanced medium, OpM1, containing growth factors and other stimulatory ingredients, which enabled the creation of the first continuous marine sponge cell line from G. barretti, achieving nearly 100 population doublings (Hesp et al. 2023). The results also demonstrated that sponge cells are capable of extremely rapid division. G. barretti cells exhibited a doubling time of under one hour in M1 and under 30 minutes in OpM1 (Hesp et al. 2023). For WP2 activities, previously successful media (Zhang et al. 2002) will serve as the starting point for new experiments aimed at achieving multiple cell passages in medium-throughput cell-culture systems using well plates, with further validation in flask-based cultures or bioreactors. The effectiveness of in vitro conditions will be assessed, based on increases in cell numbers, cell proliferation rates and culture longevity.

We further investigated the in vitro responses of various blood cell types from Botryllus (a model colonial tunicate), using five distinct formulations of cell culture media (Andrade et al. 1999). This study supports broader initiatives to develop cell type-specific cultures and highlights the critical role of tailored nutrient media in promoting cell cultures and selecting for specific cell types. For WP2 activities, this targeted approach will be further pursued and extended to coral cell culture. For echinoderms, media optimisation will involve the use of growth media supplemented with sea cucumber derived extracellular matrix (ECM) proteins and the development of an animal-component-free liquid medium suitable for both suspension and adherent cultures of sea cucumber cells. Insights gained from these efforts may also be applied to refine and optimise culture conditions for sponges, corals and ascidians.

Understanding the relationship between gene expression patterns, specifically, genes that are up- or down-regulated and the production of bioactive compounds is key to deciphering the complex biosynthetic pathways involved. This knowledge can provide information for strategies to enhance metabolite yields and clarify whether these compounds originate from the host invertebrate, its microbial symbionts or both (Munroe et al. 2019). In WP3 activities, one of the tasks is to understand the genetic mechanisms underlying both cell immortalisation and metabolite biosynthesis in marine invertebrates, as well as to explore the potential for genetic modification in these cell lines. To this end, cells will be characterised at each passage, focusing on telomerase activity and immortality markers (e.g. mortalin, survivin, klf13, Ajelp2). In addition, multi-passaged epithelial cell cultures derived from the body walls of sea cucumbers will be studied for the expression of genes associated with the biosynthesis of selected bioactive peptides, fucoidan and saponins (antimicrobial, anti-inflammatory and anti-cancer compounds with established potential) (Atanassova et al. 2024). These analyses will be conducted as part of a broader whole-expression gene ontology study. The most promising cell lines will then be tested under various culture conditions to assess: (a) changes in bioactive peptide, fucoidan and saponin synthesis and (b) shifts in immortality marker expression, allowing for the identification of optimal candidate cell lines for further development.

WP2 will also explore the development of co-culture systems of coral cell with algal symbionts as well as sponge cell with microbial symbionts. The underlying rationale is that co-cultivation with symbionts may enhance the longevity and proliferation of invertebrate cell cultures. In the case of corals, maintaining a functional association with symbiotic algae could support sustained cell growth by boosting biomass and promoting continuous cell division. These algae, which reside within the coral’s endodermal cells, provide a major portion of the host’s energy requirements through the translocation of photosynthetically fixed carbon, while the coral cells actively regulate the physiology of their symbionts, creating a mutually beneficial interaction (Radax et al. 2012).

In the case of the sponge Geodia barretti, it is believed that barettins are produced either through the symbiotic relationship between the sponge and its associated microbes or by the microbial symbionts alone (Steffen et al. 2021). After several passages of G. barretti cells, barettins were no longer detectable (partner WU, unpublished), suggesting that culture conditions, while favourable for sponge cell divisions, may have led to the loss of essential microbial symbionts. To test this hypothesis, cells from selected G. barretti passages will be analysed to determine shifts or reductions in microbial community composition, with the aim of identifying microbiome members that may be critical for sponge cell growth and/or production of barettins. Cultures will be analysed using 18S and 16S rRNA gene sequencing from different passages, alongside chemical analyses of the same samples for the presence of the barettins and the barrettides (the latter being sponge-derived compounds (Steffen et al. 2021). Based on the findings, culture conditions will be optimised, particularly through the change or removal of antibiotics in the media, to support microbiome maintenance. Further, metagenomic analyses will be performed to define potential co-metabolic pathways involved in metabolite biosynthesis.

In situations where bioactive compounds originate from species whose cells cannot be cultured or where biosynthetic pathways are unknown or difficult to engineer, hybridoma technology offers a viable alternative for developing production strains. Traditionally used to generate monoclonal antibodies by fusing antibody-producing B cells with rapidly dividing tumour cells, this approach creates cells (hybridomas) that combine high productivity with continuous cell division (Pomponi et al. 2013). A similar strategy can be applied by fusing rapidly dividing marine invertebrate cells with cells capable of producing target compounds, potentially yielding hybrid cell lines that inherit both traits (Fig. 3). We have already demonstrated the successful fusion of cells from different marine sponge species (Pomponi 2013), highlighting them as promising candidates for generating sponge-based hybridomas.

Figure 3.  

Schematic representation of the integrated process for the recovery of fucoxanthin and chlorophylls from Saccharina latissimi (Kholany et al. 2025).

As part of WP3, Geodia barretti cells will be fused with compound-producing marine invertebrate cells that currently lack the ability to divide in culture (Fig. 4). The resulting hybridomas will be screened for their ability to produce the target compound(s). Potential fusion partners include cells from corals, sea cucumbers, ascidians and other sponges, whose culture conditions are being optimised under WP2.

Figure 4.  

Illustrating sponge hybridoma principle. Uncultivable sponge cells that produce compounds of interest, simplified by the asterisks (A) can be fused with a sponge cell line from another species (B) to create a hybrid cell line (C) that inherits both traits, immortality and bioactive compound production (Cai and Zhang 2014).

Concluding Remarks

Author Contributions

Acknowledgements

Funding program

Conflicts of interest

References

Andrade P, Willoughby R, Pomponi S, Kerr R (1999)
Biosynthetic studies of the alkaloid, stevensine, in a cell culture of the marine sponge Teichaxinella morchella
.
Tetrahedron Letters
40
(
26
):
4775
‑
4778
. https://doi.org/10.1016/s0040-4039(99)00881-3
Atanassova M, Kolden Midtbo L, Mildenberger J, Friðjónsson Ó (2024)
Novel biomaterials and biotechnological applications derived from North Atlantic sea cucumbers: A systematic review
.
The World of Sea Cucumbers
585
‑
609
. https://doi.org/10.1016/b978-0-323-95377-1.00046-1
Bayne C (1998)
Chapter 10 Invertebrate Cell Culture Considerations: Insects, Ticks, Shellfish, and Worms
.
Methods in Cell Biology
187
‑
201
. https://doi.org/10.1016/s0091-679x(08)61578-2
Ben-Hamo O, Rosner A, Rabinowitz C, Oren M, Rinkevich B (2018)
Coupling astogenic aging in the colonial tunicate Botryllus schlosseri with the stress protein mortalin
.
Developmental Biology
433
(
1
):
33
‑
46
. https://doi.org/10.1016/j.ydbio.2017.10.023
Bergmann W, Feeney R (2002)
Contributions To The Study Of Marine Products. XXXII. The Nucleosides Of Sponges. I.<
.
The Journal of Organic Chemistry
16
(
6
):
981
‑
987
. https://doi.org/10.1021/jo01146a023
Brück W, Santiago-Vázquez L, Brück T (2023)
Editorial: Sustainable production of marine natural products - from discovery to application
.
Frontiers in Marine Science
10
https://doi.org/10.3389/fmars.2023.1188090
Cacciamali A, Villa R, Dotti S (2022)
3D Cell Cultures: Evolution of an Ancient Tool for New Applications
.
Frontiers in Physiology
13
https://doi.org/10.3389/fphys.2022.836480
Cai X, Zhang Y (2014)
Marine invertebrate cell culture: a decade of development
.
Journal of Oceanography
70
(
5
):
405
‑
414
. https://doi.org/10.1007/s10872-014-0242-8
Calado R, Mamede R, Cruz S, Leal M (2022)
Updated Trends on the Biodiscovery of New Marine Natural Products from Invertebrates
.
Marine Drugs
20
(
6
). https://doi.org/10.3390/md20060389
Campuzano S, Pelling A (2019)
Scaffolds for 3D Cell Culture and Cellular Agriculture Applications Derived From Non-animal Sources
.
Frontiers in Sustainable Food Systems
3
https://doi.org/10.3389/fsufs.2019.00038
Carroll A, Copp B, Davis R, Keyzers R, Prinsep M (2020)
Marine natural products
.
Natural Product Reports
37
(
2
):
175
‑
223
. https://doi.org/10.1039/c9np00069k
Carroll A, Copp B, Davis R, Keyzers R, Prinsep M (2022)
Marine natural products
.
Natural Product Reports
39
(
6
):
1122
‑
1171
. https://doi.org/10.1039/d1np00076d
Cetkovic H, Grebenjuk V, Müller WG, Gamulin V (2004)
Src proteins/src genes: from sponges to mammals
.
Gene
342
(
2
):
251
‑
261
. https://doi.org/10.1016/j.gene.2004.07.044
Cetkovic H, Mikoc A, Müller WG, Gamulin V (2007)
Ras-like Small GTPases Form a Large Family of Proteins in the Marine Sponge Suberites domuncula
.
Journal of Molecular Evolution
64
(
3
):
332
‑
341
. https://doi.org/10.1007/s00239-006-0081-3
Conkling M, Hesp K, Munroe S, Sandoval K, Martens D, Sipkema D, Wijffels R, Pomponi S (2019)
Breakthrough in Marine Invertebrate Cell Culture: Sponge Cells Divide Rapidly in Improved Nutrient Medium
.
Scientific Reports
9
(
1
). https://doi.org/10.1038/s41598-019-53643-y
Domart-Coulon I, Blanchoud S (2022)
<p>From Primary Cell and Tissue Cultures to Aquatic Invertebrate Cell Lines: An Updated Overview</p> <p>&nbsp;</p>
.
Advances in aquatic invertebrate stem cell research
https://doi.org/10.3390/books978-3-0365-1635-6-1
Duan L, Dou L, Guo L, Li P, Liu E (2016)
Comprehensive Evaluation of Deep Eutectic Solvents in Extraction of Bioactive Natural Products
.
ACS Sustainable Chemistry & Engineering
4
(
4
):
2405
‑
2411
. https://doi.org/10.1021/acssuschemeng.6b00091
Eroldoğan OT, Glencross B, Novoveska L, Gaudêncio S, Rinkevich B, Varese GC, de Fátima Carvalho M, Tasdemir D, Safarik I, Nielsen SL, Rebours C, Lada LB, Robbens J, Strode E, Haznedaroğlu B, Kotta J, Evliyaoğlu E, Oliveira J, Girão M, Vasquez M, Čabarkapa I, Rakita S, Klun K, Rotter A (2022)
From the sea to aquafeed: A perspective overview
.
Reviews in Aquaculture
15
(
3
):
1028
‑
1057
. https://doi.org/10.1111/raq.12740
Fassini D, Wilkie I, Pozzolini M, Ferrario C, Sugni M, Rocha M, Giovine M, Bonasoro F, Silva T, Reis R (2021)
Diverse and Productive Source of Biopolymer Inspiration: Marine Collagens
.
Biomacromolecules
22
(
5
):
1815
‑
1834
. https://doi.org/10.1021/acs.biomac.1c00013
Gamulin V, Rinkevich B, Schaecke H (1994)
Cell adhesion receptors and nuclear receptors are highly conserved from the lowest metazoa (marine sponges) to vertebrates
.
Biological Chemistry Hoppe-Seyler
375
(
9
):
583
‑
588
. https://doi.org/10.1515/bchm3.1994.375.9.583
Grasela J, Pomponi S, Rinkevich B, Grima J (2011)
Efforts to develop a cultured sponge cell line: revisiting an intractable problem
.
In Vitro Cellular & Developmental Biology - Animal
48
(
1
):
12
‑
20
. https://doi.org/10.1007/s11626-011-9469-5
Guryanova S, Balandin S, Belogurova-Ovchinnikova O, Ovchinnikova T (2023)
Marine Invertebrate Antimicrobial Peptides and Their Potential as Novel Peptide Antibiotics
.
Marine Drugs
21
(
10
). https://doi.org/10.3390/md21100503
Hanifaturahmah F, Dewanti-Hariyadi R, Hasanah U, Nurilmala M (2024)
Prediction and Characterization of Antimicrobial Peptides from Sea Cucumbers (Holothuria sp.) in Papuan Waters, Indonesia
.
Research Square
https://doi.org/10.21203/rs.3.rs-5430498/v1
Hesp K, van der Heijden JE, Munroe S, Sipkema D, Martens D, Wijffels R, Pomponi S (2023)
First continuous marine sponge cell line established
.
Scientific Reports
13
(
1
). https://doi.org/10.1038/s41598-023-32394-x
Hiemstra IA, Eppink MM, Bravi M, Wijffels R, Kazbar A (2025)
Process intensification of seaweed biorefinery using reusable natural deep eutectic solvents
.
Journal of Environmental Chemical Engineering
13
(
5
). https://doi.org/10.1016/j.jece.2025.119184
Hiemstra IA, Husna F, Eppink MM, Wijffels R, Kazbar A (2026)
Mechanistic insights into temperature-responsive deep eutectic solvent for alginate recovery and solvent recycling
.
Separation and Purification Technology
388
https://doi.org/10.1016/j.seppur.2026.136807
Jeon Y, Samarakoon K, Elvitigala DS (2015)
Marine-Derived Pharmaceuticals and Future Prospects
.
Hb25_Springer Handbook of Marine Biotechnology
957
‑
968
. https://doi.org/10.1007/978-3-642-53971-8_41
Kholany M, Reynaga-Navarro W, Abranches D, Wijffels R, Coutinho JP, Ventura SM, Kazbar A (2025)
Extraction and separation of pigments from Saccharina latissima using eutectic solvents
.
Separation and Purification Technology
357
https://doi.org/10.1016/j.seppur.2024.130053
Kobayashi J, Ishibashi M (2000)
Bioactive secondary metabolites from Okinawan sponges and tunicates
.
Studies in Natural Products Chemistry
185
‑
231
. https://doi.org/10.1016/s1572-5995(00)80130-1
Krasko A, Lorenz B, Batel R, Schröder H, Müller I, Müller WG (2001)
Expression of silicatein and collagen genes in the marine sponge <i>Suberites domuncula</i> is controlled by silicate and myotrophin
.
European Journal of Biochemistry
267
(
15
):
4878
‑
4887
. https://doi.org/10.1046/j.1432-1327.2000.01547.x
Li T, Ding T, Li J (2018)
Medicinal Purposes: Bioactive Metabolites from Marine-derived Organisms
.
Mini-Reviews in Medicinal Chemistry
19
(
2
):
138
‑
164
. https://doi.org/10.2174/1389557517666170927113143
Lo C, Semerel J, van den Berg C, Wijffels R, Eppink MM (2021)
Eutectic solvents with tuneable hydrophobicity: lipid dissolution and recovery
.
RSC Advances
11
(
14
):
8142
‑
8149
. https://doi.org/10.1039/d1ra00306b
Luthringer B, Isbert S, Müller WEG, Zilberberg C, Thakur NL, Wörheide G, Stauber RH, Kelve M, Wiens M (2010)
Poriferan survivin exhibits a conserved regulatory role in the interconnected pathways of cell cycle and apoptosis
.
Cell Death & Differentiation
18
(
2
):
201
‑
213
. https://doi.org/10.1038/cdd.2010.87
Ma Y, Zhang L, Huang X (2014)
Genome modification by CRISPRCas9
.
The FEBS Journal
281
(
23
):
5186
‑
5193
. https://doi.org/10.1111/febs.13110
Mothersill C, Austin B (2000)
Aquatic invertebrate cell culture
.
Aquatic invertebrate cell culture
.
409
pp. [ISBN
1852336463, 9781852336462
].
Muller W, Skorokhod A, Muller I (1999)
Receptor tyrosine kinase, an autapomorphic character of metazoa: Identification in marine sponges
.
Acta Biol Hung
50
(
4
):
395
‑
411
. https://doi.org/10.1007/BF03543061
Müller WG, Böhm M, Batel R, De Rosa S, Tommonaro G, Müller I, Schröder H (2000)
Application of Cell Culture for the Production of Bioactive Compounds from Sponges: Synthesis of Avarol by Primmorphs from <i>Dysidea</i> <i>avara</i>
.
Journal of Natural Products
63
(
8
):
1077
‑
1081
. https://doi.org/10.1021/np000003p
Munroe S, Sandoval K, Martens D, Sipkema D, Pomponi S (2019)
Genetic algorithm as an optimization tool for the development of sponge cell culture media
.
In Vitro Cellular & Developmental Biology - Animal
55
(
3
):
149
‑
158
. https://doi.org/10.1007/s11626-018-00317-0
Mutsenko V, Bazhenov V, Rogulska O, Tarusin D, Schütz K, Brüggemeier S, Gossla E, Akkineni A, Meißner H, Lode A, Meschke S, Ehrlich A, Petović S, Martinović R, Djurović M, Stelling A, Nikulin S, Rodin S, Tonevitsky A, Gelinsky M, Petrenko A, Glasmacher B, Ehrlich H (2017)
3D chitinous scaffolds derived from cultivated marine demosponge Aplysina aerophoba for tissue engineering approaches based on human mesenchymal stromal cells
.
International Journal of Biological Macromolecules
104
:
1966
‑
1974
. https://doi.org/10.1016/j.ijbiomac.2017.03.116
O'Brien F (2011)
Biomaterials & scaffolds for tissue engineering
.
Materials Today
14
(
3
):
88
‑
95
. https://doi.org/10.1016/s1369-7021(11)70058-x
Paul V, Ritson-Williams R, Sharp K (2011)
Marine chemical ecology in benthic environments
.
Nat. Prod. Rep.
28
(
2
):
345
‑
387
. https://doi.org/10.1039/c0np00040j
Pérez-López P, de Vree J, Feijoo G, Bosma R, Barbosa M, Moreira MT, Wijffels R, van Boxtel AB, Kleinegris DM (2017)
Comparative life cycle assessment of real pilot reactors for microalgae cultivation in different seasons
.
Applied Energy
205
:
1151
‑
1164
. https://doi.org/10.1016/j.apenergy.2017.08.102
Pomponi S (1997)
Developments of techniques for in vitro production of bioactive natural products from marine sponges
.
Invertebracte Cell Culture: Novel Directions and Biotechnology Application
231
‑
237
.
Pomponi S, Willoughby R, Kelly-Borges M (1998a)
Sponge Cell Culture
.
Molecular Approaches to the Study of the Ocean
423
‑
433
. https://doi.org/10.1007/978-94-011-4928-0_22
Pomponi S, Willoughby R, Wright A, Pecorella C, Sennett S, Lopez J, Samples G (1998b)
In Vitro Production of Marine-Derived Antitumor Compounds
.
New Developments in Marine Biotechnology
73
‑
76
. https://doi.org/10.1007/978-1-4757-5983-9_15
Pomponi SA (1999)
The bioprocess-technological potential of the sea
.
Progress in Industrial Microbiology
5
‑
13
. https://doi.org/10.1016/s0079-6352(99)80092-7
Pomponi SA (2006)
Biology of the Porifera: cell culture
.
Canadian Journal of Zoology
84
(
2
):
167
‑
174
. https://doi.org/10.1139/z05-188
Pomponi SA (2013)
Sponge Cell Culture State-of-the-Art: What's worked, What Hasn't, What's Next.
Cytotechnology
65
(
5
):
684
‑
689
. [In
English
]. https://doi.org/10.1007/s10616-013-9610-2
Pomponi SA, Jevitt A, Patel J, Diaz MC (2013)
Sponge Hybridomas: Applications and Implications
.
Integrative and Comparative Biology
53
(
3
):
524
‑
530
. https://doi.org/10.1093/icb/ict032
Pozzolini M, Valisano L, Cerrano C, Menta M, Schiaparelli S, Bavestrello G, Benatti U, Giovine M (2009)
Influence of rocky substrata on three-dimensional sponge cells model development
.
In Vitro Cellular & Developmental Biology - Animal
46
(
2
):
140
‑
147
. https://doi.org/10.1007/s11626-009-9253-y
Pozzolini M, Bruzzone F, Berilli V, Mussino F, Cerrano C, Benatti U, Giovine M (2011)
Molecular Characterization of a Nonfibrillar Collagen from the Marine Sponge Chondrosia reniformis Nardo 1847 and Positive Effects of Soluble Silicates on Its Expression
.
Marine Biotechnology
14
(
3
):
281
‑
293
. https://doi.org/10.1007/s10126-011-9415-2
Pozzolini M, Mussino F, Cerrano C, Scarfì S, Giovine M (2014)
Sponge cell cultivation: Optimization of the model Petrosia ficiformis (Poiret 1789)
.
Journal of Experimental Marine Biology and Ecology
454
:
70
‑
77
. https://doi.org/10.1016/j.jembe.2014.02.004
Pozzolini M, Scarfì S, Gallus L, Castellano M, Vicini S, Cortese K, Gagliani M, Bertolino M, Costa G, Giovine M (2018)
Production, Characterization and Biocompatibility Evaluation of Collagen Membranes Derived from Marine Sponge Chondrosia reniformis Nardo, 1847
.
Marine Drugs
16
(
4
). https://doi.org/10.3390/md16040111
Pozzolini M, Tassara E, Dodero A, Castellano M, Vicini S, Ferrando S, Aicardi S, Cavallo D, Bertolino M, Petrenko I, Ehrlich H, Giovine M (2021)
Potential Biomedical Applications of Collagen Filaments derived from the Marine Demosponges Ircinia oros (Schmidt, 1864) and Sarcotragus foetidus (Schmidt, 1862)
.
Marine Drugs
19
(
10
). https://doi.org/10.3390/md19100563
Puglisi M, Sneed J, Sharp K, Ritson-Williams R, Paul V (2014)
Marine chemical ecology in benthic environments
.
Nat. Prod. Rep.
31
(
11
):
1510
‑
1553
. https://doi.org/10.1039/c4np00017j
Puglisi M, Sneed J, Ritson-Williams R, Young R (2019)
Marine chemical ecology in benthic environments
.
Natural Product Reports
36
(
3
):
410
‑
429
. https://doi.org/10.1039/c8np00061a
Radax R, Rattei T, Lanzen A, Bayer C, Rapp HT, Urich T, Schleper C (2012)
Metatranscriptomics of the marine sponge<i>Geodia barretti</i>: tackling phylogeny and function of its microbial community
.
Environmental Microbiology
14
(
5
):
1308
‑
1324
. https://doi.org/10.1111/j.1462-2920.2012.02714.x
Reynaga-Navarro W, Bozzo L, Wijffels R, Eppink M, Kazbar A (2026)
Characterisation of alginate extracted from Saccharina latissima with deep eutectic solvents
.
Food Hydrocolloids
171
https://doi.org/10.1016/j.foodhyd.2025.111862
Rinkevich B (1999)
Cell cultures from marine invertebrates: obstacles, new approaches and recent improvements
.
Journal of Biotechnology
70
:
133
‑
153
. https://doi.org/10.1016/s0168-1656(99)00067-x
Rinkevich B (2005)
Marine Invertebrate Cell Cultures: New Millennium Trends
.
Marine Biotechnology
7
(
5
):
429
‑
439
. https://doi.org/10.1007/s10126-004-0108-y
Rinkevich B (2011)
Cell Cultures from Marine Invertebrates: New Insights for Capturing Endless Stemness
.
Marine Biotechnology
13
(
3
):
345
‑
354
. https://doi.org/10.1007/s10126-010-9354-3
Rodrigues IG, Miguel MG, Mnif W (2019)
A Brief Review on New Naturally Occurring Cembranoid Diterpene Derivatives from the Soft Corals of the Genera Sarcophyton, Sinularia, and Lobophytum Since 2016
.
Molecules
24
(
4
). https://doi.org/10.3390/molecules24040781
Rotter A, Barbier M, Bertoni F, Bones A, Cancela ML, Carlsson J, Carvalho M, Cegłowska M, Chirivella-Martorell J, Conk Dalay M, Cueto M, Dailianis T, Deniz I, Díaz-Marrero A, Drakulovic D, Dubnika A, Edwards C, Einarsson H, Erdoǧan A, Eroldoǧan OT, Ezra D, Fazi S, FitzGerald R, Gargan L, Gaudêncio S, Gligora Udovič M, Ivošević DeNardis N, Jónsdóttir R, Kataržytė M, Klun K, Kotta J, Ktari L, Ljubešić Z, Lukić Bilela L, Mandalakis M, Massa-Gallucci A, Matijošytė I, Mazur-Marzec H, Mehiri M, Nielsen SL, Novoveská L, Overlingė D, Perale G, Ramasamy P, Rebours C, Reinsch T, Reyes F, Rinkevich B, Robbens J, Röttinger E, Rudovica V, Sabotič J, Safarik I, Talve S, Tasdemir D, Theodotou Schneider X, Thomas O, Toruńska-Sitarz A, Varese GC, Vasquez M (2021)
The Essentials of Marine Biotechnology
.
Frontiers in Marine Science
8
https://doi.org/10.3389/fmars.2021.629629
Rotter A, Varamogianni-Mamatsi D, Zvonar Pobirk A, Gosenca Matjaž M, Cueto M, Díaz-Marrero A, Jónsdóttir R, Sveinsdóttir K, Catalá T, Romano G, Aslanbay Guler B, Atak E, Berden Zrimec M, Bosch D, Deniz I, Gaudêncio S, Grigalionyte-Bembič E, Klun K, Zidar L, Coll Rius A, Baebler Š, Lukić Bilela L, Rinkevich B, Mandalakis M (2024)
Marine cosmetics and the blue bioeconomy: From sourcing to success stories
.
iScience
27
(
12
). https://doi.org/10.1016/j.isci.2024.111339
Saeed AU, Su J, Ouyang S (2021)
Marine-derived drugs: Recent advances in cancer therapy and immune signaling
.
Biomedicine & Pharmacotherapy
134
https://doi.org/10.1016/j.biopha.2020.111091
Schäcke H, Müller WG, Gamulin V, Rinkevich B (1994)
The Ig superfamily includes members from the lowest invertebrates to the highest vertebrates
.
Immunology Today
15
(
10
):
497
‑
498
. https://doi.org/10.1016/0167-5699(94)90198-8
Sieblist C, Jenzsch M, Pohlscheidt M (2015)
Equipment characterization to mitigate risks during transfers of cell culture manufacturing processes
.
Cytotechnology
68
(
4
):
1381
‑
1401
. https://doi.org/10.1007/s10616-015-9899-0
Silva TH, Alves A, Ferreira BM, Oliveira JM, Reys LL, Ferreira RJF, Sousa RA, Silva SS, Mano JF, Reis RL (2012)
Materials of marine origin: a review on polymers and ceramics of biomedical interest
.
International Materials Reviews
57
(
5
):
276
‑
306
. https://doi.org/10.1179/1743280412y.0000000002
Sipkema D, Osinga R, Schatton W, Mendola D, Tramper J, Wijffels R (2005)
Large‐scale production of pharmaceuticals by marine sponges: Sea, cell, or synthesis?
Biotechnology and Bioengineering
90
(
2
):
201
‑
222
. https://doi.org/10.1002/bit.20404
Smith E, Abbott A, Ryder K (2014)
Deep Eutectic Solvents (DESs) and Their Applications
.
Chemical Reviews
114
(
21
):
11060
‑
11082
. https://doi.org/10.1021/cr300162p
Sorushanova A, Delgado L, Wu Z, Shologu N, Kshirsagar A, Raghunath R, Mullen A, Bayon Y, Pandit A, Raghunath M, Zeugolis D (2019)
The Collagen Suprafamily: From Biosynthesis to Advanced Biomaterial Development
.
Advanced Materials
31
(
1
). https://doi.org/10.1002/adma.201801651
Steffen K, Laborde Q, Gunasekera S, Payne C, Rosengren KJ, Riesgo A, Göransson U, Cárdenas P (2021)
Barrettides: A Peptide Family Specifically Produced by the Deep-Sea Sponge <i>Geodia barretti</i>
.
Journal of Natural Products
84
(
12
):
3138
‑
3146
. https://doi.org/10.1021/acs.jnatprod.1c00938
Swatschek D, Schatton W, Kellermann J, Müller WG, Kreuter J (2002)
Marine sponge collagen: isolation, characterization and effects on the skin parameters surface-pH, moisture and sebum
.
European Journal of Pharmaceutics and Biopharmaceutics
53
(
1
):
107
‑
113
. https://doi.org/10.1016/s0939-6411(01)00192-8
Tennakoon P, Chandika P, Yi M, Jung W (2023)
Marine-derived biopolymers as potential bioplastics, an eco-friendly alternative
.
iScience
26
(
4
). https://doi.org/10.1016/j.isci.2023.106404
Tripathi N, Shrivastava A (2019)
Recent Developments in Bioprocessing of Recombinant Proteins: Expression Hosts and Process Development
.
Frontiers in Bioengineering and Biotechnology
7
https://doi.org/10.3389/fbioe.2019.00420
Tsurkan D, Wysokowski M, Petrenko I, Voronkina A, Khrunyk Y, Fursov A, Ehrlich H (2020)
Modern scaffolding strategies based on naturally pre-fabricated 3D biomaterials of poriferan origin
.
Applied Physics A
126
(
5
). https://doi.org/10.1007/s00339-020-03564-9
United Nations (2018)
UN Environment Program, Convention on Biological Diversity
. https://www.cbd.int/article/biodiversityforwater-1. Accessed on: 2018-11-26.
Wang X, Schröder H, Müller WG (2014)
Enzyme-based biosilica and biocalcite: biomaterials for the future in regenerative medicine
.
Trends in Biotechnology
32
(
9
):
441
‑
447
. https://doi.org/10.1016/j.tibtech.2014.05.004
Wiens M, Krasko A, Müller C, Müller WG (2000)
Molecular Evolution of Apoptotic Pathways: Cloning of Key Domains from Sponges (Bcl-2 Homology Domains and Death Domains) and Their Phylogenetic Relationships
.
Journal of Molecular Evolution
50
(
6
):
520
‑
531
. https://doi.org/10.1007/s002390010055
Willoughby R, Pomponi S (2000)
Quantitative Assment Of MarineSponge Cells In Vitro: Develpoment Of Improved Growth Medium
.
In Vitro Cellular & Developmental Biology - Animal
36
(
3
). https://doi.org/10.1290/1071-2690(2000)0362.0.co;2
Wung N, Acott S, Tosh D, Ellis M (2014)
Hollow fibre membrane bioreactors for tissue engineering applications
.
Biotechnology Letters
36
(
12
):
2357
‑
2366
. https://doi.org/10.1007/s10529-014-1619-x
Yeshanew M, Frunzo L, Luongo V, Pirozzi F, Lens PL, Esposito G (2016)
Start-up of an anaerobic fluidized bed reactor treating synthetic carbohydrate rich wastewater
.
Journal of Environmental Management
184
:
456
‑
464
. https://doi.org/10.1016/j.jenvman.2016.10.001
Zhang X, Zhao Q, Xue S, Zhang W (2002)
[Bioactive compounds from marine sponges and cell culture of marine sponges].
Sheng wu gong cheng xue bao = Chinese journal of biotechnology
18
(
1
):
10
‑
5
.
Zhao Y, Hao L, Meng Y, Li L, Wang W, Zhao R, Zhao P, Zhang J, Wang M, Ren J, Zhang L, Yin X, Xia X (2024)
Screening and heterologous expression of an antimicrobial peptide SCAK33 with broad-spectrum antimicrobial activity resourced from sea cucumber proteome
.
International Microbiology
28
(
5
):
1019
‑
1031
. https://doi.org/10.1007/s10123-024-00595-7

Supplementary material