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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
Online Access: Check full text

URI: https://cronfa.swan.ac.uk/Record/cronfa51993
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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. 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.
Keywords: Bone marrow, blunt trauma, hemorrhagic shock (HS), flow Cytometry, hematopoietic progenitor cells (HPC), granulocytes, monocytes, lymphocytes
College: Faculty of Medicine, Health and Life Sciences
Issue: 11
Start Page: 1167
End Page: 1177

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