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Flow Cytometric Analysis of Hematopoietic Populations in Rat Bone Marrow. Impact of Trauma and Hemorrhagic Shock

Wendy Francis Orcid Logo, Rachel E. Ireland, Abigail M. Spear, Dominic Jenner, Sarah A. Watts, Emrys Kirkman, Ian Pallister

Cytometry Part A, Volume: 95, Issue: 11, Pages: 1167 - 1177

Swansea University Authors: Wendy Francis Orcid Logo, Ian Pallister

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DOI (Published version): 10.1002/cyto.a.23903

Abstract

Severe injury and hemorrhagic shock (HS) result in multiple changes to hematopoietic differentiation, which contribute to the development of immunosuppression and multiple organ failure (MOF). Understanding the changes that take place during the acute injury phase may help predict which patients wil...

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Published in: Cytometry Part A
ISSN: 1552-4922 1552-4930
Published: Wiley 2019
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Impact of Trauma and Hemorrhagic Shock</title><swanseaauthors><author><sid>f0ec2a3fdae1cf112579d579afbe9813</sid><ORCID>0000-0002-7952-2770</ORCID><firstname>Wendy</firstname><surname>Francis</surname><name>Wendy Francis</name><active>true</active><ethesisStudent>false</ethesisStudent></author><author><sid>8ddd73971cbb1d06f5ee3a0ce8a90e58</sid><firstname>Ian</firstname><surname>Pallister</surname><name>Ian Pallister</name><active>true</active><ethesisStudent>false</ethesisStudent></author></swanseaauthors><date>2019-09-23</date><deptcode>BMS</deptcode><abstract>Severe injury and hemorrhagic shock (HS) result in multiple changes to hematopoietic differentiation, which contribute to the development of immunosuppression and multiple organ failure (MOF). Understanding the changes that take place during the acute injury phase may help predict which patients will develop MOF and provide potential targets for therapy. Obtaining bone marrow from humans during the acute injury phase is difficult so published data is largely derived from peripheral blood samples, which infer bone marrow changes that reflect the sustained inflammatory response. This preliminary and opportunistic study investigated leucopoietic changes in rat bone marrow 6 hours following traumatic injury and HS. Terminally anesthetized male Porton Wistar rats were allocated randomly to receive a sham operation (cannulation with no injury) or femoral fracture and HS. Bone marrow cells were flushed from rat femurs and immunophenotypically stained with specific antibody panels for lymphoid (CD45R, CD127, CD90, IgM) or myeloid (CD11b, CD45, RP-1) lineages. Subsequently, cell populations were fluorescence activated cell sorted for morphological assessment. Stage-specific cell populations were identified using a limited number of antibodies and leucopoietic changes were determined 6 hours following trauma and HS. Myeloid sub-populations could be identified by varying levels CD11b expression, CD45 and RP-1. Trauma and HS resulted in a significant reduction in total CD11b+ myeloid cells including both immature (RP-1(-)) and mature (RP-1+) granulocytes. Multiple B-cell lymphoid subsets were identified. 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spelling 2019-10-08T12:38:49.4398458 v2 51993 2019-09-23 Flow Cytometric Analysis of Hematopoietic Populations in Rat Bone Marrow. Impact of Trauma and Hemorrhagic Shock f0ec2a3fdae1cf112579d579afbe9813 0000-0002-7952-2770 Wendy Francis Wendy Francis true false 8ddd73971cbb1d06f5ee3a0ce8a90e58 Ian Pallister Ian Pallister true false 2019-09-23 BMS Severe injury and hemorrhagic shock (HS) result in multiple changes to hematopoietic differentiation, which contribute to the development of immunosuppression and multiple organ failure (MOF). Understanding the changes that take place during the acute injury phase may help predict which patients will develop MOF and provide potential targets for therapy. Obtaining bone marrow from humans during the acute injury phase is difficult so published data is largely derived from peripheral blood samples, which infer bone marrow changes that reflect the sustained inflammatory response. This preliminary and opportunistic study investigated leucopoietic changes in rat bone marrow 6 hours following traumatic injury and HS. Terminally anesthetized male Porton Wistar rats were allocated randomly to receive a sham operation (cannulation with no injury) or femoral fracture and HS. Bone marrow cells were flushed from rat femurs and immunophenotypically stained with specific antibody panels for lymphoid (CD45R, CD127, CD90, IgM) or myeloid (CD11b, CD45, RP-1) lineages. Subsequently, cell populations were fluorescence activated cell sorted for morphological assessment. Stage-specific cell populations were identified using a limited number of antibodies and leucopoietic changes were determined 6 hours following trauma and HS. Myeloid sub-populations could be identified by varying levels CD11b expression, CD45 and RP-1. Trauma and HS resulted in a significant reduction in total CD11b+ myeloid cells including both immature (RP-1(-)) and mature (RP-1+) granulocytes. Multiple B-cell lymphoid subsets were identified. The total % of CD90+ subsets remained unchanged following trauma and HS, but there was a reduction in the numbers of maturing CD90(-) cells suggesting movement into the periphery. Journal Article Cytometry Part A 95 11 1167 1177 Wiley 1552-4922 1552-4930 Bone marrow, blunt trauma, hemorrhagic shock (HS), flow Cytometry, hematopoietic progenitor cells (HPC), granulocytes, monocytes, lymphocytes 30 11 2019 2019-11-30 10.1002/cyto.a.23903 COLLEGE NANME Biomedical Sciences COLLEGE CODE BMS Swansea University MoD CSA Research Programme 2019-10-08T12:38:49.4398458 2019-09-23T09:31:41.3920754 Faculty of Medicine, Health and Life Sciences Swansea University Medical School - Medicine Wendy Francis 0000-0002-7952-2770 1 Rachel E. Ireland 2 Abigail M. Spear 3 Dominic Jenner 4 Sarah A. Watts 5 Emrys Kirkman 6 Ian Pallister 7 51993__15930__af59c50690df46a9a433ce3403d0532d.pdf 51993.pdf 2019-11-21T10:14:35.2890979 Output 3962690 application/pdf Version of Record true Released under the terms of a Creative Commons Attribution-NonCommercial License (CC-BY-NC). true https://creativecommons.org/licenses/by-nc/4.0/
title Flow Cytometric Analysis of Hematopoietic Populations in Rat Bone Marrow. Impact of Trauma and Hemorrhagic Shock
spellingShingle Flow Cytometric Analysis of Hematopoietic Populations in Rat Bone Marrow. Impact of Trauma and Hemorrhagic Shock
Wendy Francis
Ian Pallister
title_short Flow Cytometric Analysis of Hematopoietic Populations in Rat Bone Marrow. Impact of Trauma and Hemorrhagic Shock
title_full Flow Cytometric Analysis of Hematopoietic Populations in Rat Bone Marrow. Impact of Trauma and Hemorrhagic Shock
title_fullStr Flow Cytometric Analysis of Hematopoietic Populations in Rat Bone Marrow. Impact of Trauma and Hemorrhagic Shock
title_full_unstemmed Flow Cytometric Analysis of Hematopoietic Populations in Rat Bone Marrow. Impact of Trauma and Hemorrhagic Shock
title_sort Flow Cytometric Analysis of Hematopoietic Populations in Rat Bone Marrow. Impact of Trauma and Hemorrhagic Shock
author_id_str_mv f0ec2a3fdae1cf112579d579afbe9813
8ddd73971cbb1d06f5ee3a0ce8a90e58
author_id_fullname_str_mv f0ec2a3fdae1cf112579d579afbe9813_***_Wendy Francis
8ddd73971cbb1d06f5ee3a0ce8a90e58_***_Ian Pallister
author Wendy Francis
Ian Pallister
author2 Wendy Francis
Rachel E. Ireland
Abigail M. Spear
Dominic Jenner
Sarah A. Watts
Emrys Kirkman
Ian Pallister
format Journal article
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container_volume 95
container_issue 11
container_start_page 1167
publishDate 2019
institution Swansea University
issn 1552-4922
1552-4930
doi_str_mv 10.1002/cyto.a.23903
publisher Wiley
college_str Faculty of Medicine, Health and Life Sciences
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hierarchy_top_title Faculty of Medicine, Health and Life Sciences
hierarchy_parent_id facultyofmedicinehealthandlifesciences
hierarchy_parent_title Faculty of Medicine, Health and Life Sciences
department_str Swansea University Medical School - Medicine{{{_:::_}}}Faculty of Medicine, Health and Life Sciences{{{_:::_}}}Swansea University Medical School - Medicine
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description Severe injury and hemorrhagic shock (HS) result in multiple changes to hematopoietic differentiation, which contribute to the development of immunosuppression and multiple organ failure (MOF). Understanding the changes that take place during the acute injury phase may help predict which patients will develop MOF and provide potential targets for therapy. Obtaining bone marrow from humans during the acute injury phase is difficult so published data is largely derived from peripheral blood samples, which infer bone marrow changes that reflect the sustained inflammatory response. This preliminary and opportunistic study investigated leucopoietic changes in rat bone marrow 6 hours following traumatic injury and HS. Terminally anesthetized male Porton Wistar rats were allocated randomly to receive a sham operation (cannulation with no injury) or femoral fracture and HS. Bone marrow cells were flushed from rat femurs and immunophenotypically stained with specific antibody panels for lymphoid (CD45R, CD127, CD90, IgM) or myeloid (CD11b, CD45, RP-1) lineages. Subsequently, cell populations were fluorescence activated cell sorted for morphological assessment. Stage-specific cell populations were identified using a limited number of antibodies and leucopoietic changes were determined 6 hours following trauma and HS. Myeloid sub-populations could be identified by varying levels CD11b expression, CD45 and RP-1. Trauma and HS resulted in a significant reduction in total CD11b+ myeloid cells including both immature (RP-1(-)) and mature (RP-1+) granulocytes. Multiple B-cell lymphoid subsets were identified. The total % of CD90+ subsets remained unchanged following trauma and HS, but there was a reduction in the numbers of maturing CD90(-) cells suggesting movement into the periphery.
published_date 2019-11-30T04:04:06Z
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