Paralytic Impact of Centrifugation on Human Neutrophils
Centrifugation is a common step in most of the popular protocols for the isolation of neutrophils from whole blood. Inconsistent results from previous studies on neutrophils may originate from an underestimation of the centrifugation effect, as in consequence impaired, not native cells, being invest...
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| Format: | Article |
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MDPI AG
2022-11-01
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| Series: | Biomedicines |
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| Online Access: | https://www.mdpi.com/2227-9059/10/11/2896 |
| _version_ | 1797465865777577984 |
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| author | Tobias Hundhammer Michael Gruber Sigrid Wittmann |
| author_facet | Tobias Hundhammer Michael Gruber Sigrid Wittmann |
| author_sort | Tobias Hundhammer |
| collection | DOAJ |
| description | Centrifugation is a common step in most of the popular protocols for the isolation of neutrophils from whole blood. Inconsistent results from previous studies on neutrophils may originate from an underestimation of the centrifugation effect, as in consequence impaired, not native cells, being investigated. We hypothesize, that centrifugation significantly impairs major neutrophil functions. However, there is no data yet whether the application of <i>g</i>-force itself or the product of <i>g</i>-force and duration of centrifugation (=“<i>g</i>-time”) defines the impact on neutrophils. Neutrophils were isolated from whole blood via centrifugation with different <i>g</i>-times and subsequently analyzed via live cell imaging for migration, as well as via flow cytometry for oxidative burst and surface antigen expression. Chemotactic migration was significantly reduced with increasing <i>g</i>-time. Oxidative burst decreased likewise the higher the <i>g</i>-time applied. Expression of CD11b was no longer upregulated in response to an n-formylmethionine-leucyl-phenylalanine (fMLP) stimulus in neutrophils having experienced high <i>g</i>-time during the isolation process. We conclude that centrifugation “paralyzes” neutrophils in the form of a significant decrease in functionality. Future investigations on neutrophil granulocytes should reduce the <i>g</i>-time load as far as possible. |
| first_indexed | 2024-03-09T18:27:39Z |
| format | Article |
| id | doaj.art-ad5bfd0c82f24cb79cc95c0fb09efc9e |
| institution | Directory Open Access Journal |
| issn | 2227-9059 |
| language | English |
| last_indexed | 2024-03-09T18:27:39Z |
| publishDate | 2022-11-01 |
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| series | Biomedicines |
| spelling | doaj.art-ad5bfd0c82f24cb79cc95c0fb09efc9e2023-11-24T07:45:50ZengMDPI AGBiomedicines2227-90592022-11-011011289610.3390/biomedicines10112896Paralytic Impact of Centrifugation on Human NeutrophilsTobias Hundhammer0Michael Gruber1Sigrid Wittmann2Department of Anesthesiology, University Hospital Regensburg, 93042 Regensburg, GermanyDepartment of Anesthesiology, University Hospital Regensburg, 93042 Regensburg, GermanyDepartment of Anesthesiology, University Hospital Regensburg, 93042 Regensburg, GermanyCentrifugation is a common step in most of the popular protocols for the isolation of neutrophils from whole blood. Inconsistent results from previous studies on neutrophils may originate from an underestimation of the centrifugation effect, as in consequence impaired, not native cells, being investigated. We hypothesize, that centrifugation significantly impairs major neutrophil functions. However, there is no data yet whether the application of <i>g</i>-force itself or the product of <i>g</i>-force and duration of centrifugation (=“<i>g</i>-time”) defines the impact on neutrophils. Neutrophils were isolated from whole blood via centrifugation with different <i>g</i>-times and subsequently analyzed via live cell imaging for migration, as well as via flow cytometry for oxidative burst and surface antigen expression. Chemotactic migration was significantly reduced with increasing <i>g</i>-time. Oxidative burst decreased likewise the higher the <i>g</i>-time applied. Expression of CD11b was no longer upregulated in response to an n-formylmethionine-leucyl-phenylalanine (fMLP) stimulus in neutrophils having experienced high <i>g</i>-time during the isolation process. We conclude that centrifugation “paralyzes” neutrophils in the form of a significant decrease in functionality. Future investigations on neutrophil granulocytes should reduce the <i>g</i>-time load as far as possible.https://www.mdpi.com/2227-9059/10/11/2896neutrophilsPMNsisolationcentrifugation<i>g</i>-time<i>g</i>-forces |
| spellingShingle | Tobias Hundhammer Michael Gruber Sigrid Wittmann Paralytic Impact of Centrifugation on Human Neutrophils Biomedicines neutrophils PMNs isolation centrifugation <i>g</i>-time <i>g</i>-forces |
| title | Paralytic Impact of Centrifugation on Human Neutrophils |
| title_full | Paralytic Impact of Centrifugation on Human Neutrophils |
| title_fullStr | Paralytic Impact of Centrifugation on Human Neutrophils |
| title_full_unstemmed | Paralytic Impact of Centrifugation on Human Neutrophils |
| title_short | Paralytic Impact of Centrifugation on Human Neutrophils |
| title_sort | paralytic impact of centrifugation on human neutrophils |
| topic | neutrophils PMNs isolation centrifugation <i>g</i>-time <i>g</i>-forces |
| url | https://www.mdpi.com/2227-9059/10/11/2896 |
| work_keys_str_mv | AT tobiashundhammer paralyticimpactofcentrifugationonhumanneutrophils AT michaelgruber paralyticimpactofcentrifugationonhumanneutrophils AT sigridwittmann paralyticimpactofcentrifugationonhumanneutrophils |