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Cnidarian Cell Cryopreservation: A Powerful Tool for Cultivation and Functional Assays

Cnidarian primary cell cultures have a strong potential to become a universal tool to assess stress-response mechanisms at the cellular level. However, primary cell cultures are time-consuming regarding their establishment and maintenance. Cryopreservation is a commonly used approach to provide stab...

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Autores principales: Fricano, Clara, Röttinger, Eric, Furla, Paola, Barnay-Verdier, Stéphanie
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7761476/
https://www.ncbi.nlm.nih.gov/pubmed/33256018
http://dx.doi.org/10.3390/cells9122541
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author Fricano, Clara
Röttinger, Eric
Furla, Paola
Barnay-Verdier, Stéphanie
author_facet Fricano, Clara
Röttinger, Eric
Furla, Paola
Barnay-Verdier, Stéphanie
author_sort Fricano, Clara
collection PubMed
description Cnidarian primary cell cultures have a strong potential to become a universal tool to assess stress-response mechanisms at the cellular level. However, primary cell cultures are time-consuming regarding their establishment and maintenance. Cryopreservation is a commonly used approach to provide stable cell stocks for experiments, but it is yet to be established for Cnidarian cell cultures. The aim of this study was therefore to design a cryopreservation protocol for primary cell cultures of the Cnidarian Anemonia viridis, using dimethyl sulfoxide (DMSO) as a cryoprotectant, enriched or not with fetal bovine serum (FBS). We determined that DMSO 5% with 25% FBS was an efficient cryosolution, resulting in 70% of post-thaw cell survival. The success of this protocol was first confirmed by a constant post-thaw survival independently of the cell culture age (up to 45 days old) and the storage period (up to 87 days). Finally, cryopreserved cells displayed a long-term recovery with a maintenance of the primary cell culture parameters and cellular functions: formation of cell aggregates, high viability and constant cell growth, and unchanged intrinsic resistance to hyperthermal stress. These results will further bring new opportunities for the scientific community interested in molecular, cellular, and biochemical aspects of cnidarian biology.
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spelling pubmed-77614762020-12-26 Cnidarian Cell Cryopreservation: A Powerful Tool for Cultivation and Functional Assays Fricano, Clara Röttinger, Eric Furla, Paola Barnay-Verdier, Stéphanie Cells Article Cnidarian primary cell cultures have a strong potential to become a universal tool to assess stress-response mechanisms at the cellular level. However, primary cell cultures are time-consuming regarding their establishment and maintenance. Cryopreservation is a commonly used approach to provide stable cell stocks for experiments, but it is yet to be established for Cnidarian cell cultures. The aim of this study was therefore to design a cryopreservation protocol for primary cell cultures of the Cnidarian Anemonia viridis, using dimethyl sulfoxide (DMSO) as a cryoprotectant, enriched or not with fetal bovine serum (FBS). We determined that DMSO 5% with 25% FBS was an efficient cryosolution, resulting in 70% of post-thaw cell survival. The success of this protocol was first confirmed by a constant post-thaw survival independently of the cell culture age (up to 45 days old) and the storage period (up to 87 days). Finally, cryopreserved cells displayed a long-term recovery with a maintenance of the primary cell culture parameters and cellular functions: formation of cell aggregates, high viability and constant cell growth, and unchanged intrinsic resistance to hyperthermal stress. These results will further bring new opportunities for the scientific community interested in molecular, cellular, and biochemical aspects of cnidarian biology. MDPI 2020-11-26 /pmc/articles/PMC7761476/ /pubmed/33256018 http://dx.doi.org/10.3390/cells9122541 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Fricano, Clara
Röttinger, Eric
Furla, Paola
Barnay-Verdier, Stéphanie
Cnidarian Cell Cryopreservation: A Powerful Tool for Cultivation and Functional Assays
title Cnidarian Cell Cryopreservation: A Powerful Tool for Cultivation and Functional Assays
title_full Cnidarian Cell Cryopreservation: A Powerful Tool for Cultivation and Functional Assays
title_fullStr Cnidarian Cell Cryopreservation: A Powerful Tool for Cultivation and Functional Assays
title_full_unstemmed Cnidarian Cell Cryopreservation: A Powerful Tool for Cultivation and Functional Assays
title_short Cnidarian Cell Cryopreservation: A Powerful Tool for Cultivation and Functional Assays
title_sort cnidarian cell cryopreservation: a powerful tool for cultivation and functional assays
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7761476/
https://www.ncbi.nlm.nih.gov/pubmed/33256018
http://dx.doi.org/10.3390/cells9122541
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