Development of separation and characterisation techniques for cells of the marine sponge Hymeniacidon perlevis

Details

Supervisors

Rees, Jean-François

Faculty

Faculté des bioingénieurs

Degree label

Master [120] : bioingénieur en sciences agronomiques, à finalité spécialisée

Abstract

In recent years, marine sponges have attracted the interest of scientists and industry for their biotechnological potential. These seemingly simple animals hide a surprising complexity. On the one hand, their structural plasticity and the totipotency of their cells have attracted the interest of regenerative medicine. On the other hand, sponges produce a variety of chemical compounds with anticancer, antimicrobial, or antifouling properties that can address health or environmental concerns. However, the development of new spongederived technologies is severely hampered by the inability to obtain sufficient sponge biomass. Recently, new perspectives in cell culture have emerged and some specific sponge cell types (namely archaeocytes and choanocytes) have been identified as having higher growth potential than others. This project is part of a larger effort to grow sponges using 3D cell printing as a biomass production method. The globally distributed marine sponge Hymeniacidon perlevis is the first candidate for the development of a 3D culture model. To better understand the biology of Hymeniacidon perlevis, the cells and structure of this species were characterised using optical and scanning electron microscopy. The choanocytes and archaeocytes were clearly identified and several hypotheses regarding the arrangement of the extracellular matrix were formulated. A method of separating cells by density using a Percoll gradient was successfully developed in an attempt to isolate cell types, in particular choanocytes and archaeocytes. The fractions from the gradient were characterised optically and using a flow cytometer, which demonstrated the effectiveness of the gradient in enriching fractions according to cell size and complexity. Based on visual observations, the gradient also appears to have allowed enrichment of archaeocytes and choanocytes. This work has shown that density gradient, flow cytometry as well as optical and scanning electron imaging techniques are effective in separating, analysing, and imaging sponge cells. These methods can be used for cell culture of fractions enriched in specific cell types. In addition, the characterisation of the structure of Hymeniacidon perlevis provides a key to the future development of a structural model for 3D printing. In the long term, 3D printing of sponges could provide the first bioprinted animal model and serve as a basis for the development of new tissue printing techniques or the production of new sponge-derived drugs.