Project

Immortalizing marine sponge cell culture

Marine sponge cells are the richest natural source of biologically active and/or commercially attractive compounds. The goal of project is to obtain stable and/or immortalized sponge cell lines required to harness this enormous potential through the use of novel biotechnological techniques such as RNA interference and CRISPR-Cas9 gene editing.

Introduction

Since harvesting sponges for bioactive compounds is neither environmentally nor economically feasible, an alternative strategy for the bulk production of sponge based bioactive compounds is necessary. Large-scale in vitro sponge cell culture may provide a well-defined and controllable environment for the production of chemicals and other bioproducts of interest. However, due to significant gaps in our understanding of culture conditions for marine invertebrate cells, all attempts to maintain a normal or immortalized marine invertebrate cell line have been unsuccessful.

Aim and research question

The aim of this project is to develop immortalized sponge cell cultures for the production of marine- derived bioproducts.

Approach

The first step involves the optimization of primary sponge cell cultures. By measuring the cell cycle distribution and apoptosis we can get more insight in the proliferative status ofsponge cells and we can compare different cultivation techniques, such as tissue culture vs. suspension cells.

Next to this, we are investigating the possibilities of expressing heterologous genes in juvenile freshwater sponges. We are comparing different gene delivery methods, such aslipofection and particle bombardment. The next step will be the transfection of juvenile sponges with reporter genes, such as GFP and luciferase. Finally, we will transfect sponge cells with immortalizing genes and translate these results to marine species.

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Acknowledgements

This research is supported by IPOP Coast and Sea, Graduate School VLAG and the Florida Sea Grant (R/LR-MB-25) and is in collaboration with Harbor Branch Oceanographic Institute, USA.