Coral cell separation and isolation by fluorescence-activated cell sorting (FACS)
Abstract Background Generalized methods for understanding the cell biology of non-model species are quite rare, yet very much needed. In order to address this issue, we have modified a technique traditionally used in the biomedical field for ecological and evolutionary research. Fluorescent activate...
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| Format: | Article |
| Language: | English |
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BMC
2017-08-01
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| Series: | BMC Cell Biology |
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| Online Access: | http://link.springer.com/article/10.1186/s12860-017-0146-8 |
| _version_ | 1811235875805200384 |
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| author | Benyamin Rosental Zhanna Kozhekbaeva Nathaniel Fernhoff Jonathan M. Tsai Nikki Traylor-Knowles |
| author_facet | Benyamin Rosental Zhanna Kozhekbaeva Nathaniel Fernhoff Jonathan M. Tsai Nikki Traylor-Knowles |
| author_sort | Benyamin Rosental |
| collection | DOAJ |
| description | Abstract Background Generalized methods for understanding the cell biology of non-model species are quite rare, yet very much needed. In order to address this issue, we have modified a technique traditionally used in the biomedical field for ecological and evolutionary research. Fluorescent activated cell sorting (FACS) is often used for sorting and identifying cell populations. In this study, we developed a method to identify and isolate different cell populations in corals and other cnidarians. Methods Using fluorescence-activated cell sorting (FACS), coral cell suspension were sorted into different cellular populations using fluorescent cell markers that are non-species specific. Over 30 different cell markers were tested. Additionally, cell suspension from Aiptasia pallida was also tested, and a phagocytosis test was done as a downstream functional assay. Results We found that 24 of the screened markers positively labeled coral cells and 16 differentiated cell sub-populations. We identified 12 different cellular sub-populations using three markers, and found that each sub-population is primarily homogeneous. Lastly, we verified this technique in a sea anemone, Aiptasia pallida, and found that with minor modifications, a similar gating strategy can be successfully applied. Additionally, within A. pallida, we show elevated phagocytosis of sorted cells based on an immune associated marker. Conclusions In this study, we successfully adapted FACS for isolating coral cell populations and conclude that this technique is translatable for future use in other species. This technique has the potential to be used for different types of studies on the cellular stress response and other immunological studies. |
| first_indexed | 2024-04-12T11:59:37Z |
| format | Article |
| id | doaj.art-da0d4f7f4f054027b8d19e98a12afff0 |
| institution | Directory Open Access Journal |
| issn | 1471-2121 |
| language | English |
| last_indexed | 2024-04-12T11:59:37Z |
| publishDate | 2017-08-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Cell Biology |
| spelling | doaj.art-da0d4f7f4f054027b8d19e98a12afff02022-12-22T03:33:55ZengBMCBMC Cell Biology1471-21212017-08-0118111210.1186/s12860-017-0146-8Coral cell separation and isolation by fluorescence-activated cell sorting (FACS)Benyamin Rosental0Zhanna Kozhekbaeva1Nathaniel Fernhoff2Jonathan M. Tsai3Nikki Traylor-Knowles4Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of MedicineUniversity of Miami, Rosenstiel School of Marine and Atmospheric ScienceInstitute for Stem Cell Biology and Regenerative Medicine, Stanford University School of MedicineInstitute for Stem Cell Biology and Regenerative Medicine, Stanford University School of MedicineUniversity of Miami, Rosenstiel School of Marine and Atmospheric ScienceAbstract Background Generalized methods for understanding the cell biology of non-model species are quite rare, yet very much needed. In order to address this issue, we have modified a technique traditionally used in the biomedical field for ecological and evolutionary research. Fluorescent activated cell sorting (FACS) is often used for sorting and identifying cell populations. In this study, we developed a method to identify and isolate different cell populations in corals and other cnidarians. Methods Using fluorescence-activated cell sorting (FACS), coral cell suspension were sorted into different cellular populations using fluorescent cell markers that are non-species specific. Over 30 different cell markers were tested. Additionally, cell suspension from Aiptasia pallida was also tested, and a phagocytosis test was done as a downstream functional assay. Results We found that 24 of the screened markers positively labeled coral cells and 16 differentiated cell sub-populations. We identified 12 different cellular sub-populations using three markers, and found that each sub-population is primarily homogeneous. Lastly, we verified this technique in a sea anemone, Aiptasia pallida, and found that with minor modifications, a similar gating strategy can be successfully applied. Additionally, within A. pallida, we show elevated phagocytosis of sorted cells based on an immune associated marker. Conclusions In this study, we successfully adapted FACS for isolating coral cell populations and conclude that this technique is translatable for future use in other species. This technique has the potential to be used for different types of studies on the cellular stress response and other immunological studies.http://link.springer.com/article/10.1186/s12860-017-0146-8FACSCoralCell isolationCell sortingCellular labeling |
| spellingShingle | Benyamin Rosental Zhanna Kozhekbaeva Nathaniel Fernhoff Jonathan M. Tsai Nikki Traylor-Knowles Coral cell separation and isolation by fluorescence-activated cell sorting (FACS) BMC Cell Biology FACS Coral Cell isolation Cell sorting Cellular labeling |
| title | Coral cell separation and isolation by fluorescence-activated cell sorting (FACS) |
| title_full | Coral cell separation and isolation by fluorescence-activated cell sorting (FACS) |
| title_fullStr | Coral cell separation and isolation by fluorescence-activated cell sorting (FACS) |
| title_full_unstemmed | Coral cell separation and isolation by fluorescence-activated cell sorting (FACS) |
| title_short | Coral cell separation and isolation by fluorescence-activated cell sorting (FACS) |
| title_sort | coral cell separation and isolation by fluorescence activated cell sorting facs |
| topic | FACS Coral Cell isolation Cell sorting Cellular labeling |
| url | http://link.springer.com/article/10.1186/s12860-017-0146-8 |
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