An evaluation of sorter induced cell stress (SICS) on peripheral blood mononuclear cells (PBMCs) after different sort conditions - Are your sorted cells getting SICS?
<p dir="ltr">Flow cytometry and fluorescence-activated cell sorting have become invaluable tools to analyze and isolate specific cell populations in a wide range of biomedical research and clinical applications. In countless approaches worldwide, scientists are using single cell anal...
محفوظ في:
| المؤلف الرئيسي: | |
|---|---|
| مؤلفون آخرون: | , , |
| منشور في: |
2020
|
| الموضوعات: | |
| الوسوم: |
إضافة وسم
لا توجد وسوم, كن أول من يضع وسما على هذه التسجيلة!
|
| _version_ | 1864513559063429120 |
|---|---|
| author | Gerald Pfister (17075275) |
| author2 | Salman M. Toor (8854751) Varun Sasidharan Nair (5396393) Eyad Elkord (5396390) |
| author2_role | author author author |
| author_facet | Gerald Pfister (17075275) Salman M. Toor (8854751) Varun Sasidharan Nair (5396393) Eyad Elkord (5396390) |
| author_role | author |
| dc.creator.none.fl_str_mv | Gerald Pfister (17075275) Salman M. Toor (8854751) Varun Sasidharan Nair (5396393) Eyad Elkord (5396390) |
| dc.date.none.fl_str_mv | 2020-12-01T00:00:00Z |
| dc.identifier.none.fl_str_mv | 10.1016/j.jim.2020.112902 |
| dc.relation.none.fl_str_mv | https://figshare.com/articles/journal_contribution/An_evaluation_of_sorter_induced_cell_stress_SICS_on_peripheral_blood_mononuclear_cells_PBMCs_after_different_sort_conditions_-_Are_your_sorted_cells_getting_SICS_/24225664 |
| dc.rights.none.fl_str_mv | CC BY 4.0 info:eu-repo/semantics/openAccess |
| dc.subject.none.fl_str_mv | Biomedical and clinical sciences Immunology Cell sorting Flow cytometry FACS Cell purification Sorter induced cell stress SICS Viability Proliferation T cells PBMC CFSE 7AAD |
| dc.title.none.fl_str_mv | An evaluation of sorter induced cell stress (SICS) on peripheral blood mononuclear cells (PBMCs) after different sort conditions - Are your sorted cells getting SICS? |
| dc.type.none.fl_str_mv | Text Journal contribution info:eu-repo/semantics/publishedVersion text contribution to journal |
| description | <p dir="ltr">Flow cytometry and fluorescence-activated cell sorting have become invaluable tools to analyze and isolate specific cell populations in a wide range of biomedical research and clinical applications. In countless approaches worldwide, scientists are using single cell analyses to better understand the significance and variation within different cellular populations, and fluorescence-activated cell sorting has become a major technique for cell isolation in both basic and clinical research. However, majority of available cell sorters are pressurized, droplet-based systems, which apply significant environmental pressure and shear stress to cells during sorting. Recently, the flow cytometry community has become increasingly aware about the potential negative effects this could have on sorted cells and the term “sorter induced cell stress” (SICS) has been proposed. However, up to date only a limited number of studies have investigated the effects of cell sorting on cell viability and function. Therefore, solid data on the effects of sheath pressure and nozzle size on survival and function of sorted cells are surprisingly rare. With this in mind, we sorted “CD4+” T-cells and “live” cells from human peripheral blood mononuclear cells (PBMCs) at different sort conditions and analyzed their quality before and after sorting in a series of assays. Here we present our findings in reference to cell viability and cell proliferation following sorting on different instruments (BD FACSAria III SORP and BD FACSJazz), utilizing different nozzle sizes (70 to 100 μm) and sheath pressure settings (20 to 70 psi). The results show no significant differences in cell viability and proliferation after the different tested sort conditions, but rather differences between individual experiments. These findings are evaluated and their potential significance in cell sorting experiments is discussed.</p><h2>Other Information</h2><p dir="ltr">Published in: Journal of Immunological Methods<br>License: <a href="http://creativecommons.org/licenses/by/4.0/" target="_blank">http://creativecommons.org/licenses/by/4.0/</a><br>See article on publisher's website: <a href="https://dx.doi.org/10.1016/j.jim.2020.112902" target="_blank">https://dx.doi.org/10.1016/j.jim.2020.112902</a></p> |
| eu_rights_str_mv | openAccess |
| id | Manara2_47b299de6629c0be09cfbd28e0f8b899 |
| identifier_str_mv | 10.1016/j.jim.2020.112902 |
| network_acronym_str | Manara2 |
| network_name_str | Manara2 |
| oai_identifier_str | oai:figshare.com:article/24225664 |
| publishDate | 2020 |
| repository.mail.fl_str_mv | |
| repository.name.fl_str_mv | |
| repository_id_str | |
| rights_invalid_str_mv | CC BY 4.0 |
| spelling | An evaluation of sorter induced cell stress (SICS) on peripheral blood mononuclear cells (PBMCs) after different sort conditions - Are your sorted cells getting SICS?Gerald Pfister (17075275)Salman M. Toor (8854751)Varun Sasidharan Nair (5396393)Eyad Elkord (5396390)Biomedical and clinical sciencesImmunologyCell sortingFlow cytometryFACSCell purificationSorter induced cell stressSICSViabilityProliferationT cellsPBMCCFSE7AAD<p dir="ltr">Flow cytometry and fluorescence-activated cell sorting have become invaluable tools to analyze and isolate specific cell populations in a wide range of biomedical research and clinical applications. In countless approaches worldwide, scientists are using single cell analyses to better understand the significance and variation within different cellular populations, and fluorescence-activated cell sorting has become a major technique for cell isolation in both basic and clinical research. However, majority of available cell sorters are pressurized, droplet-based systems, which apply significant environmental pressure and shear stress to cells during sorting. Recently, the flow cytometry community has become increasingly aware about the potential negative effects this could have on sorted cells and the term “sorter induced cell stress” (SICS) has been proposed. However, up to date only a limited number of studies have investigated the effects of cell sorting on cell viability and function. Therefore, solid data on the effects of sheath pressure and nozzle size on survival and function of sorted cells are surprisingly rare. With this in mind, we sorted “CD4+” T-cells and “live” cells from human peripheral blood mononuclear cells (PBMCs) at different sort conditions and analyzed their quality before and after sorting in a series of assays. Here we present our findings in reference to cell viability and cell proliferation following sorting on different instruments (BD FACSAria III SORP and BD FACSJazz), utilizing different nozzle sizes (70 to 100 μm) and sheath pressure settings (20 to 70 psi). The results show no significant differences in cell viability and proliferation after the different tested sort conditions, but rather differences between individual experiments. These findings are evaluated and their potential significance in cell sorting experiments is discussed.</p><h2>Other Information</h2><p dir="ltr">Published in: Journal of Immunological Methods<br>License: <a href="http://creativecommons.org/licenses/by/4.0/" target="_blank">http://creativecommons.org/licenses/by/4.0/</a><br>See article on publisher's website: <a href="https://dx.doi.org/10.1016/j.jim.2020.112902" target="_blank">https://dx.doi.org/10.1016/j.jim.2020.112902</a></p>2020-12-01T00:00:00ZTextJournal contributioninfo:eu-repo/semantics/publishedVersiontextcontribution to journal10.1016/j.jim.2020.112902https://figshare.com/articles/journal_contribution/An_evaluation_of_sorter_induced_cell_stress_SICS_on_peripheral_blood_mononuclear_cells_PBMCs_after_different_sort_conditions_-_Are_your_sorted_cells_getting_SICS_/24225664CC BY 4.0info:eu-repo/semantics/openAccessoai:figshare.com:article/242256642020-12-01T00:00:00Z |
| spellingShingle | An evaluation of sorter induced cell stress (SICS) on peripheral blood mononuclear cells (PBMCs) after different sort conditions - Are your sorted cells getting SICS? Gerald Pfister (17075275) Biomedical and clinical sciences Immunology Cell sorting Flow cytometry FACS Cell purification Sorter induced cell stress SICS Viability Proliferation T cells PBMC CFSE 7AAD |
| status_str | publishedVersion |
| title | An evaluation of sorter induced cell stress (SICS) on peripheral blood mononuclear cells (PBMCs) after different sort conditions - Are your sorted cells getting SICS? |
| title_full | An evaluation of sorter induced cell stress (SICS) on peripheral blood mononuclear cells (PBMCs) after different sort conditions - Are your sorted cells getting SICS? |
| title_fullStr | An evaluation of sorter induced cell stress (SICS) on peripheral blood mononuclear cells (PBMCs) after different sort conditions - Are your sorted cells getting SICS? |
| title_full_unstemmed | An evaluation of sorter induced cell stress (SICS) on peripheral blood mononuclear cells (PBMCs) after different sort conditions - Are your sorted cells getting SICS? |
| title_short | An evaluation of sorter induced cell stress (SICS) on peripheral blood mononuclear cells (PBMCs) after different sort conditions - Are your sorted cells getting SICS? |
| title_sort | An evaluation of sorter induced cell stress (SICS) on peripheral blood mononuclear cells (PBMCs) after different sort conditions - Are your sorted cells getting SICS? |
| topic | Biomedical and clinical sciences Immunology Cell sorting Flow cytometry FACS Cell purification Sorter induced cell stress SICS Viability Proliferation T cells PBMC CFSE 7AAD |