Establishing Cell Lines from Fresh or Cryopreserved Tissue from the Great Crested Newt (<i>Triturus cristatus</i>): A Preliminary Protocol
This study describes a successful protocol for establishing cell lines from the threatened <i>Triturus cristatus</i> in terms of collection, preparing, establishing, cryopreserving, thawing and quality checking. Different parameters such as media, media change, fresh vs. cryopreserved ti...
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| Format: | Article |
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MDPI AG
2021-02-01
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| Series: | Animals |
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| Online Access: | https://www.mdpi.com/2076-2615/11/2/367 |
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| author | Julie Strand Henrik Callesen Cino Pertoldi Stig Purup |
| author_facet | Julie Strand Henrik Callesen Cino Pertoldi Stig Purup |
| author_sort | Julie Strand |
| collection | DOAJ |
| description | This study describes a successful protocol for establishing cell lines from the threatened <i>Triturus cristatus</i> in terms of collection, preparing, establishing, cryopreserving, thawing and quality checking. Different parameters such as media, media change, fresh vs. cryopreserved tissue and seeding density were tested to optimize culture conditions for this species. With fresh tissue, no considerable differences in the use of two different media were found, but with cryopreserved tissue, a combination of ITS (insulin/transferrin/selenite) and 2-mercaptoethanol had a positive effect on growth. Real-time measurements on the cell lines were used, for the first time in amphibian cells, to investigate the effect of different treatments such as media change with or without washing. Media change had a positive impact on the cells, whereas the effect was negative when combined with washing. It is concluded that establishment of cell lines is possible from the great crested newt, especially when using fresh tissue, but much more challenging if the tissue has been cryopreserved. Real-time measurement during cell culture is a useful tool to visualize the sensitivity of amphibian cells during different culture treatments. |
| first_indexed | 2024-03-09T06:09:21Z |
| format | Article |
| id | doaj.art-6b418b308bc44d969c64c7ab2aa2a00c |
| institution | Directory Open Access Journal |
| issn | 2076-2615 |
| language | English |
| last_indexed | 2024-03-09T06:09:21Z |
| publishDate | 2021-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Animals |
| spelling | doaj.art-6b418b308bc44d969c64c7ab2aa2a00c2023-12-03T12:00:15ZengMDPI AGAnimals2076-26152021-02-0111236710.3390/ani11020367Establishing Cell Lines from Fresh or Cryopreserved Tissue from the Great Crested Newt (<i>Triturus cristatus</i>): A Preliminary ProtocolJulie Strand0Henrik Callesen1Cino Pertoldi2Stig Purup3Department of Chemistry and Bioscience, Aalborg University, Fredrik Bajers Vej 7K, DK-9220 Aalborg Øst, DenmarkDepartment of Animal Science, Aarhus University, Blichers Allé 20, DK-8830 Tjele, DenmarkDepartment of Chemistry and Bioscience, Aalborg University, Fredrik Bajers Vej 7K, DK-9220 Aalborg Øst, DenmarkDepartment of Animal Science, Aarhus University, Blichers Allé 20, DK-8830 Tjele, DenmarkThis study describes a successful protocol for establishing cell lines from the threatened <i>Triturus cristatus</i> in terms of collection, preparing, establishing, cryopreserving, thawing and quality checking. Different parameters such as media, media change, fresh vs. cryopreserved tissue and seeding density were tested to optimize culture conditions for this species. With fresh tissue, no considerable differences in the use of two different media were found, but with cryopreserved tissue, a combination of ITS (insulin/transferrin/selenite) and 2-mercaptoethanol had a positive effect on growth. Real-time measurements on the cell lines were used, for the first time in amphibian cells, to investigate the effect of different treatments such as media change with or without washing. Media change had a positive impact on the cells, whereas the effect was negative when combined with washing. It is concluded that establishment of cell lines is possible from the great crested newt, especially when using fresh tissue, but much more challenging if the tissue has been cryopreserved. Real-time measurement during cell culture is a useful tool to visualize the sensitivity of amphibian cells during different culture treatments.https://www.mdpi.com/2076-2615/11/2/367cell culturemorphologyculture conditionsxCELLigence<sup>®</sup>real-time cell analysisamphibian |
| spellingShingle | Julie Strand Henrik Callesen Cino Pertoldi Stig Purup Establishing Cell Lines from Fresh or Cryopreserved Tissue from the Great Crested Newt (<i>Triturus cristatus</i>): A Preliminary Protocol Animals cell culture morphology culture conditions xCELLigence<sup>®</sup> real-time cell analysis amphibian |
| title | Establishing Cell Lines from Fresh or Cryopreserved Tissue from the Great Crested Newt (<i>Triturus cristatus</i>): A Preliminary Protocol |
| title_full | Establishing Cell Lines from Fresh or Cryopreserved Tissue from the Great Crested Newt (<i>Triturus cristatus</i>): A Preliminary Protocol |
| title_fullStr | Establishing Cell Lines from Fresh or Cryopreserved Tissue from the Great Crested Newt (<i>Triturus cristatus</i>): A Preliminary Protocol |
| title_full_unstemmed | Establishing Cell Lines from Fresh or Cryopreserved Tissue from the Great Crested Newt (<i>Triturus cristatus</i>): A Preliminary Protocol |
| title_short | Establishing Cell Lines from Fresh or Cryopreserved Tissue from the Great Crested Newt (<i>Triturus cristatus</i>): A Preliminary Protocol |
| title_sort | establishing cell lines from fresh or cryopreserved tissue from the great crested newt i triturus cristatus i a preliminary protocol |
| topic | cell culture morphology culture conditions xCELLigence<sup>®</sup> real-time cell analysis amphibian |
| url | https://www.mdpi.com/2076-2615/11/2/367 |
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