Journal article Open Access

Breakthrough in marine invertebrate cell culture: Sponge cells divide rapidly in improved nutrient medium

Conkling, Megan; Hesp, Kylie; Munroe, Stephanie; Sandoval, Kenneth; Martens, Dirk E; Sipkema, Detmer; Wijffels, Rene; Pomponi, Shirley A

ABSTRACT.

Sponges (phylum porifera) are among the oldest Metazoa and considered critical to understanding animal evolution and development. They are also the most prolific source of marine-derived chemicals with pharmaceutical relevance. Cell lines are important tools for research in many disciplines, and have been established for many organisms, including freshwater and terrestrial invertebrates. Despite many efforts over multiple decades, there are still no cell lines for marine invertebrates. In this study, we report a breakthrough: we demonstrate that an amino acid-optimized nutrient medium stimulates rapid cell division in 9 sponge species. The fastest dividing cells doubled in less than 1 hour. Cultures of 3 species were subcultured from 3 to 5 times, with an average of 5.99 population doublings after subculturing, and a lifespan from 21 to 35 days. Our results form the basis for developing marine invertebrate cell models to better understand early animal evolution, determine the role of secondary metabolites, and predict the impact of climate change to coral reef community ecology. Furthermore, sponge cell lines can be used to scale-up production of sponge-derived chemicals for clinical trials and develop new drugs to combat cancer and other diseases.

ACKNOWLEDGMENTS: This research was supported by the European Union Marie Curie Grant (ITN-2013-BluePharmTrain-607786) (to D.S.), the European Union Horizon 2020 Project SponGES (grant agreement No. 679848) (to D.S., S.P., D.M.), the Harbor Branch Oceanographic Institute Foundation, Aquaculture and Save Our Seas Specialty License Program (to S.P.), and the National Oceanic and Atmospheric Administration, Cooperative Institute for Ocean Exploration, Research, and Technology (award number NA14OAR43202600 (to S.P.). This document reflects only the authors' views; sponsors are not responsible for any use that may be made of the information it contains. We thank Dr. M. Cristina Diaz (HBOI-FAU) and Dr. Patricia Blackwelder (Nova Southeastern University) for microscopic analyses and discussions regarding the color change in nutrient media. We thank Dr. Hans Tore Rapp (University of Bergen) for collecting Geodia barretti samples. S.P. also acknowledges the many collaborators, undergraduate and graduate students (especially Dr. Robin Willoughby and Dr. Klaske Schippers), post-doctoral fellows, and research assistants whose contributions to the sponge biotechnology programs at Florida Atlantic University-Harbor Branch Oceanographic Institute and Wageningen University provided the foundation for the success achieved in this study. AUTHOR CONTRIBUTION. M.C. conceived, designed, and conducted experiments for all species except G. barretti, analyzed data for all species except G. barretti, assisted with preparation of figures for the paper, and wrote sections of the paper. K.H. conceived, designed, and conducted experiments for G. barretti, analyzed data for G. barretti, prepared figures for the paper, and wrote sections of the paper. S.M. and K.S. conceived, designed, and conducted experiments for nutrient media optimization and the development of medium M1. D.M. conceived and designed experiments, analyzed data, contributed reagents/materials/analysis tools, and reviewed drafts of the paper. D.S. conceived and designed experiments, analyzed data, contributed reagents/materials/analysis tools, wrote sections of the paper, and reviewed drafts of the paper. R.W. conceived and designed experiments, contributed reagents/materials/ analysis tools, and reviewed drafts of the paper. S.P. conceived, designed, and conducted experiments, collected the sponges, supervised the research, analyzed data, contributed reagents/materials/analysis tools and field expenses, and wrote sections of the paper. CORRESPONDENCE. Correspondence and requests for materials should be addressed to S.A.P. COPYRIGHT. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Cre- ative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons license and your intended use is not per- mitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. © The Author(s) 2019
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