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...
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2020-11-01
|
| Series: | Cells |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2073-4409/9/12/2541 |
| _version_ | 1797546718189846528 |
|---|---|
| author | Clara Fricano Eric Röttinger Paola Furla Stéphanie Barnay-Verdier |
| author_facet | Clara Fricano Eric Röttinger Paola Furla Stéphanie Barnay-Verdier |
| author_sort | Clara Fricano |
| collection | DOAJ |
| 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 <i>Anemonia viridis</i>, 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. |
| first_indexed | 2024-03-10T14:33:13Z |
| format | Article |
| id | doaj.art-a4b5aee29f6948e3a8d5acf1b4a8ee58 |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-10T14:33:13Z |
| publishDate | 2020-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-a4b5aee29f6948e3a8d5acf1b4a8ee582023-11-20T22:25:23ZengMDPI AGCells2073-44092020-11-01912254110.3390/cells9122541Cnidarian Cell Cryopreservation: A Powerful Tool for Cultivation and Functional AssaysClara Fricano0Eric Röttinger1Paola Furla2Stéphanie Barnay-Verdier3CNRS, INSERM, Institute for Research on Cancer and Aging (IRCAN), Université Côte d’Azur, 28 avenue de Valombrose, F-06107 Nice, FranceCNRS, INSERM, Institute for Research on Cancer and Aging (IRCAN), Université Côte d’Azur, 28 avenue de Valombrose, F-06107 Nice, FranceCNRS, INSERM, Institute for Research on Cancer and Aging (IRCAN), Université Côte d’Azur, 28 avenue de Valombrose, F-06107 Nice, FranceCNRS, INSERM, Institute for Research on Cancer and Aging (IRCAN), Université Côte d’Azur, 28 avenue de Valombrose, F-06107 Nice, FranceCnidarian 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 <i>Anemonia viridis</i>, 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.https://www.mdpi.com/2073-4409/9/12/2541primary cell culturesea anemone<i>Anemonia viridis</i>dimethyl sulfoxide (DMSO)marine invertebratepost-thaw recovery |
| spellingShingle | Clara Fricano Eric Röttinger Paola Furla Stéphanie Barnay-Verdier Cnidarian Cell Cryopreservation: A Powerful Tool for Cultivation and Functional Assays Cells primary cell culture sea anemone <i>Anemonia viridis</i> dimethyl sulfoxide (DMSO) marine invertebrate post-thaw recovery |
| 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 | primary cell culture sea anemone <i>Anemonia viridis</i> dimethyl sulfoxide (DMSO) marine invertebrate post-thaw recovery |
| url | https://www.mdpi.com/2073-4409/9/12/2541 |
| work_keys_str_mv | AT clarafricano cnidariancellcryopreservationapowerfultoolforcultivationandfunctionalassays AT ericrottinger cnidariancellcryopreservationapowerfultoolforcultivationandfunctionalassays AT paolafurla cnidariancellcryopreservationapowerfultoolforcultivationandfunctionalassays AT stephaniebarnayverdier cnidariancellcryopreservationapowerfultoolforcultivationandfunctionalassays |