Protocol for the analysis of hematopoietic lineages in the whole kidney marrow of adult zebrafish

Chris Mahony; Rui Monteiro

doi:10.1016/j.xpro.2023.102810

Protocol for the analysis of hematopoietic lineages in the whole kidney marrow of adult zebrafish

Chris Mahony, Rui Monteiro^*

^*Corresponding author for this work

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Abstract

The whole kidney marrow (WKM) is the site for hematopoiesis in the adult zebrafish. Here, we present a protocol for analyzing hematopoietic lineages in the WKM of adult zebrafish. We describe steps for the isolation of hematopoietic cells from the WKM, the downstream analysis of total marrow cellularity, and analysis of cell populations by flow cytometry. We then detail procedures for May-Grünwald-Giemsa staining for analysis of cellular morphology and phenotyping.

For complete details on the use and execution of this protocol, please refer to Mahony et al.

Original language	English
Article number	102810
Number of pages	12
Journal	STAR Protocols
Volume	5
Issue number	1
Early online date	22 Jan 2024
DOIs	https://doi.org/10.1016/j.xpro.2023.102810
Publication status	Published - 15 Mar 2024

Bibliographical note

Acknowledgments
We are grateful to the staff of the Biomedical Services Unit for fish husbandry and the staff at the Flow Cytometry Facility and Dr. Adriana Flores-Langarica, Dr. Paola Pietroni, and Dr. Guillaume Desanti for cell sorting. This research was supported by a Wellcome Trust ISSF Award to R.M., a British Heart Foundation Fellowship to R.M. and C.B.M. (BHF IBSR FS/13/50/30436), and a British Heart Foundation Project Grant to R.M. (PG/22/11161).

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10.1016/j.xpro.2023.102810Licence: Creative Commons: Attribution (CC BY)

MahonyC2023ProtocolFinal published version, 5.24 MBLicence: Creative Commons: Attribution (CC BY)

The role of epigenetic regulation of haemato-endothelial cell differentiation by class IIa histone deacetylases
Monteiro, R. (Principal Investigator)
British Heart Foundation
1/05/23 → 30/04/26
Project: Research

Cite this

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abstract = "The whole kidney marrow (WKM) is the site for hematopoiesis in the adult zebrafish. Here, we present a protocol for analyzing hematopoietic lineages in the WKM of adult zebrafish. We describe steps for the isolation of hematopoietic cells from the WKM, the downstream analysis of total marrow cellularity, and analysis of cell populations by flow cytometry. We then detail procedures for May-Gr{\"u}nwald-Giemsa staining for analysis of cellular morphology and phenotyping.For complete details on the use and execution of this protocol, please refer to Mahony et al.",

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Protocol for the analysis of hematopoietic lineages in the whole kidney marrow of adult zebrafish

Abstract

Bibliographical note

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The role of epigenetic regulation of haemato-endothelial cell differentiation by class IIa histone deacetylases

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