S Munroe1, D E Martens2, D Sipkema3, S A Pomponi4. 1. Wageningen University & Research, Bioprocess Engineering (Droevendaalsesteeg 1, 6708 PB Wageningen, The Netherlands). Harbor Branch Oceanographic Institute, Florida Atlantic University (5600 US 1N, Fort Pierce, FL 34946, USA). stephanie.munroe@wur.nl. 2. Wageningen University & Research, Bioprocess Engineering (Droevendaalsesteeg 1, 6708 PB Wageningen, The Netherlands). 3. Wageningen University & Research, Laboratory of Microbiology (Stippeneng 4, 6708 WE Wageningen, The Netherlands). 4. Wageningen University & Research, Bioprocess Engineering (Droevendaalsesteeg 1, 6708 PB Wageningen, The Netherlands). Harbor Branch Oceanographic Institute, Florida Atlantic University (5600 US 1N, Fort Pierce, FL 34946, USA).
Abstract
BACKGROUND: Cryopreservation is a commonly used method for the long-term storage of cell lines and provides a stable source of cells for experiments, allowing researchers to study species that are not geographically nearby, and useful to progress studies on sponge cell biotechnology. OBJECTIVE: The marine sponge Dysidea etheria was chosen as our model organism to evaluate the impact and effectiveness of two commonly used cryoprotectants, dimethyl sulfoxide (DMSO) and glycerol. MATERIALS AND METHODS: By testing a range of concentrations (3-10% DMSO, 10-50% glycerol), we determined the optimal cryoprotectant for D. etheria based on its ability to preserve viable cells and optimize recovery after cryopreservation. RESULTS: Cells cryopreserved in DMSO had significantly higher viability after cryopreservation than those cryopreserved in glycerol. Cells cryopreserved in glycerol had irregular morphology as well as lower recovery of viable cells than those from DMSO treatments. CONCLUSION: Our results demonstrate that the optimal cryoprotectant for sponge cells, without a significant loss of viability, is 5-8% DMSO. This approach can be used to optimize cryopreservation methods for cells of other marine invertebrate species.
BACKGROUND: Cryopreservation is a commonly used method for the long-term storage of cell lines and provides a stable source of cells for experiments, allowing researchers to study species that are not geographically nearby, and useful to progress studies on sponge cell biotechnology. OBJECTIVE: The marine sponge Dysidea etheria was chosen as our model organism to evaluate the impact and effectiveness of two commonly used cryoprotectants, dimethyl sulfoxide (DMSO) and glycerol. MATERIALS AND METHODS: By testing a range of concentrations (3-10% DMSO, 10-50% glycerol), we determined the optimal cryoprotectant for D. etheria based on its ability to preserve viable cells and optimize recovery after cryopreservation. RESULTS: Cells cryopreserved in DMSO had significantly higher viability after cryopreservation than those cryopreserved in glycerol. Cells cryopreserved in glycerol had irregular morphology as well as lower recovery of viable cells than those from DMSO treatments. CONCLUSION: Our results demonstrate that the optimal cryoprotectant for sponge cells, without a significant loss of viability, is 5-8% DMSO. This approach can be used to optimize cryopreservation methods for cells of other marine invertebrate species.
Authors: Anak Agung Gede Indraningrat; Sebastian Micheller; Mandy Runderkamp; Ina Sauerland; Leontine E Becking; Hauke Smidt; Detmer Sipkema Journal: Mar Drugs Date: 2019-10-11 Impact factor: 5.118
Authors: Elizabeth Urban-Gedamke; Megan Conkling; Peter J McCarthy; Paul S Wills; Shirley A Pomponi Journal: Mar Drugs Date: 2021-10-14 Impact factor: 5.118