Limitations in the isolation and stimulation of splenic mononuclear cells in a dasyurid marsupial, Phascogale calura
Abstract Objective Marsupials suffer from an increasing number of stressors in this changing world. Functional studies are thus needed to broaden our understanding of the marsupial immune system. The red-tailed phascogale (Phascogale calura) is a small Australian marsupial previously used in descrip...
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| Format: | Article |
| Language: | English |
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BMC
2018-10-01
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| Series: | BMC Research Notes |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13104-018-3824-5 |
| _version_ | 1811194393116278784 |
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| author | C. Letendre L. J. Young J. M. Old |
| author_facet | C. Letendre L. J. Young J. M. Old |
| author_sort | C. Letendre |
| collection | DOAJ |
| description | Abstract Objective Marsupials suffer from an increasing number of stressors in this changing world. Functional studies are thus needed to broaden our understanding of the marsupial immune system. The red-tailed phascogale (Phascogale calura) is a small Australian marsupial previously used in descriptive immunological studies. Here, we aimed to develop functional assays by isolating and stimulating blood and spleen mononuclear cells in vitro. Results While peripheral blood mononuclear cell (PBMC) were relatively easy to isolate, only 105 mononuclear cells (> 90% purity and > 75% viability) could be recovered from the spleen, independently of the sex and age of the animal or the centrifugation time and speed tested. The pores of the mesh sieve used for tissue homogenization might have been too big to yield a single cell suspension. Nevertheless, in spite of the overall low number of cells recovered, PBMC and splenic mononuclear cells were successfully activated in preliminary trials with phytohemaglutinin. This activation state was evidenced by a change in shape and the presence of small cell aggregations in the mitogen-stimulated cultures. A non-radioactive colorimetric assay was also performed to confirm cell proliferation in these wells. This work highlights the importance of developing and reporting detailed methodological protocols in non-traditional research species. |
| first_indexed | 2024-04-12T00:26:22Z |
| format | Article |
| id | doaj.art-3f65d23c0de342f9a8be2e3c44514021 |
| institution | Directory Open Access Journal |
| issn | 1756-0500 |
| language | English |
| last_indexed | 2024-04-12T00:26:22Z |
| publishDate | 2018-10-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Research Notes |
| spelling | doaj.art-3f65d23c0de342f9a8be2e3c445140212022-12-22T03:55:29ZengBMCBMC Research Notes1756-05002018-10-011111710.1186/s13104-018-3824-5Limitations in the isolation and stimulation of splenic mononuclear cells in a dasyurid marsupial, Phascogale caluraC. Letendre0L. J. Young1J. M. Old2School of Science and Health, Western Sydney UniversitySchool of Science and Health, Western Sydney UniversitySchool of Science and Health, Western Sydney UniversityAbstract Objective Marsupials suffer from an increasing number of stressors in this changing world. Functional studies are thus needed to broaden our understanding of the marsupial immune system. The red-tailed phascogale (Phascogale calura) is a small Australian marsupial previously used in descriptive immunological studies. Here, we aimed to develop functional assays by isolating and stimulating blood and spleen mononuclear cells in vitro. Results While peripheral blood mononuclear cell (PBMC) were relatively easy to isolate, only 105 mononuclear cells (> 90% purity and > 75% viability) could be recovered from the spleen, independently of the sex and age of the animal or the centrifugation time and speed tested. The pores of the mesh sieve used for tissue homogenization might have been too big to yield a single cell suspension. Nevertheless, in spite of the overall low number of cells recovered, PBMC and splenic mononuclear cells were successfully activated in preliminary trials with phytohemaglutinin. This activation state was evidenced by a change in shape and the presence of small cell aggregations in the mitogen-stimulated cultures. A non-radioactive colorimetric assay was also performed to confirm cell proliferation in these wells. This work highlights the importance of developing and reporting detailed methodological protocols in non-traditional research species.http://link.springer.com/article/10.1186/s13104-018-3824-5MarsupialsDasyuridSpleenPBMCMononuclear cellsCell culture |
| spellingShingle | C. Letendre L. J. Young J. M. Old Limitations in the isolation and stimulation of splenic mononuclear cells in a dasyurid marsupial, Phascogale calura BMC Research Notes Marsupials Dasyurid Spleen PBMC Mononuclear cells Cell culture |
| title | Limitations in the isolation and stimulation of splenic mononuclear cells in a dasyurid marsupial, Phascogale calura |
| title_full | Limitations in the isolation and stimulation of splenic mononuclear cells in a dasyurid marsupial, Phascogale calura |
| title_fullStr | Limitations in the isolation and stimulation of splenic mononuclear cells in a dasyurid marsupial, Phascogale calura |
| title_full_unstemmed | Limitations in the isolation and stimulation of splenic mononuclear cells in a dasyurid marsupial, Phascogale calura |
| title_short | Limitations in the isolation and stimulation of splenic mononuclear cells in a dasyurid marsupial, Phascogale calura |
| title_sort | limitations in the isolation and stimulation of splenic mononuclear cells in a dasyurid marsupial phascogale calura |
| topic | Marsupials Dasyurid Spleen PBMC Mononuclear cells Cell culture |
| url | http://link.springer.com/article/10.1186/s13104-018-3824-5 |
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