Development of a protocol for assessing cell viability and proliferation of marine sponge cells in alginate hydrogel in the context of FRESH 3D bioprinting

Details

Supervisors

Rees, Jean-François

Faculty

Faculté des sciences

Degree label

Master [120] en biochimie et biologie moléculaire et cellulaire, à finalité spécialisée: biotechnologie

Abstract

enSponges are considered as one of the oldest multicellular non-vertebrate animals on Earth and colonized the benthic environment 600 million years ago. These primitive animals do not have complex structures but are of significant pharmaceutical interest because sponges and their associated symbiotic microorganisms produce a large quantity of bioactive compounds which have antibacterial, antibiotic, anti-cancer or anti-inflammatory effects and that can be used in human medicine. Sponges presents also important ecological interest as they are filter-feeders and thus can filter and degrade organic matter, which is interesting for the bioremediation of wastewater produced by the intensive aquaculture field. However, the population of sponges are limited in the environment and the wild harvesting of this organism for study proposal is ecologically not sustainable. Thus the need to increase sponge biomass for applied and fundamental studies is becoming increasingly crucial. Thanks to their ability to transdifferentiate, sponges are promising candidates for 3D bioprinting. The present master’s thesis is a study taking place in a large project of 3D bioprinting of sponge tissue held in the laboratory of Professor Jean-François Rees in UCLouvain. The aim of this study is to provide a complete new protocol to assess sponge cell viability and proliferation in an alginate hydrogel that will be used as starting material for the bioink in the context of FRESH 3D bioprinting technology. To develop the protocol, we first needed to develop an AH suitable for cell culture and microscopic observations. We tested 2 different methods for that and choose the best one for the development of the protocol. Then, we developed a proliferation assay based on the measurement of sponge cell pigments absorbance and we have optimised the MTT test protocol to make it compatible with measurements of metabolic activity in sponge cells cultured in AH. We performed these 2 assays in either non dissolved AH and in dissolved AH using sodium citrate and compared the difference absorbance values. We observed that the absorbance values were much higher regardless of the test and conditions when AH was dissolved compared to when it was not. Based on the different results obtained, we developed and provide an optimized protocol for the assessment of sponge cell viability and proliferation in an AH. This protocol can be used in future experiments to determine if AH is suitable and biocompatible with sponge cells for 3D bioprinting purposes.