Quantifying the Dynamics of Hematopoiesis by In Vivo IdU Pulse-Chase, Mass Cytometry, and Mathematical Modeling

Amir Erez; Ratnadeep Mukherjee; Grégoire Altan-Bonnet

doi:10.1002/cyto.a.23799

Quantifying the Dynamics of Hematopoiesis by In Vivo IdU Pulse-Chase, Mass Cytometry, and Mathematical Modeling

Amir Erez, Ratnadeep Mukherjee, Grégoire Altan-Bonnet^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

Abstract

We present a new method to directly quantify the dynamics of differentiation of multiple cellular subsets in unperturbed mice. We combine a pulse-chase protocol of 5-iodo-2′-deoxyuridine (IdU) injections with subsequent analysis by mass cytometry (CyTOF) and mathematical modeling of the IdU dynamics. Measurements by CyTOF allow for a wide range of cells to be analyzed at once, due to the availability of a large staining panel without the complication of fluorescence spillover. These are also compatible with direct detection of integrated iodine signal, with minimal impact on immunophenotyping based on the surface markers. Mathematical modeling beyond a binary classification of surface marker abundance allows for a continuum of cellular states as the cells transition from one state to another. Thus, we present a complete and robust method for directly quantifying differentiation at the systemic level, allowing for system-wide comparisons between different mouse strains and/or experimental conditions. Published 2019. This article is a U.S. Government work and is in the public domain in the USA.

Original language	American English
Pages (from-to)	1075-1084
Number of pages	10
Journal	Cytometry. Part A : the journal of the International Society for Analytical Cytology
Volume	95
Issue number	10
DOIs	https://doi.org/10.1002/cyto.a.23799
State	Published - 1 Oct 2019
Externally published	Yes

Bibliographical note

Funding Information:
This work was supported by Human Frontier Science Program grant LT000123/2014 (Amir Erez), the Gordon Moore foundation and by the Intramural Research Program of the NCI, NIH. We thank April Huang for her help with tagging the mice. We thank Brian Sellers for expert advice with mass cytometry. The CyTOF Facility at the Center for Human Immunology is supported by NIH intramural funding (AI001226-01).

Publisher Copyright:
Published 2019. This article is a U.S. Government work and is in the public domain in the USA.

Keywords

B-cells
CyTOF
IdU
bone marrow
differentiation
hematopoiesis
neutrophils

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10.1002/cyto.a.23799

Cite this

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title = "Quantifying the Dynamics of Hematopoiesis by In Vivo IdU Pulse-Chase, Mass Cytometry, and Mathematical Modeling",

abstract = "We present a new method to directly quantify the dynamics of differentiation of multiple cellular subsets in unperturbed mice. We combine a pulse-chase protocol of 5-iodo-2′-deoxyuridine (IdU) injections with subsequent analysis by mass cytometry (CyTOF) and mathematical modeling of the IdU dynamics. Measurements by CyTOF allow for a wide range of cells to be analyzed at once, due to the availability of a large staining panel without the complication of fluorescence spillover. These are also compatible with direct detection of integrated iodine signal, with minimal impact on immunophenotyping based on the surface markers. Mathematical modeling beyond a binary classification of surface marker abundance allows for a continuum of cellular states as the cells transition from one state to another. Thus, we present a complete and robust method for directly quantifying differentiation at the systemic level, allowing for system-wide comparisons between different mouse strains and/or experimental conditions. Published 2019. This article is a U.S. Government work and is in the public domain in the USA.",

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author = "Amir Erez and Ratnadeep Mukherjee and Gr{\'e}goire Altan-Bonnet",

note = "Funding Information: This work was supported by Human Frontier Science Program grant LT000123/2014 (Amir Erez), the Gordon Moore foundation and by the Intramural Research Program of the NCI, NIH. We thank April Huang for her help with tagging the mice. We thank Brian Sellers for expert advice with mass cytometry. The CyTOF Facility at the Center for Human Immunology is supported by NIH intramural funding (AI001226-01). Publisher Copyright: Published 2019. This article is a U.S. Government work and is in the public domain in the USA.",

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Quantifying the Dynamics of Hematopoiesis by In Vivo IdU Pulse-Chase, Mass Cytometry, and Mathematical Modeling. / Erez, Amir; Mukherjee, Ratnadeep; Altan-Bonnet, Grégoire.
In: Cytometry. Part A : the journal of the International Society for Analytical Cytology, Vol. 95, No. 10, 01.10.2019, p. 1075-1084.

Research output: Contribution to journal › Article › peer-review

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N2 - We present a new method to directly quantify the dynamics of differentiation of multiple cellular subsets in unperturbed mice. We combine a pulse-chase protocol of 5-iodo-2′-deoxyuridine (IdU) injections with subsequent analysis by mass cytometry (CyTOF) and mathematical modeling of the IdU dynamics. Measurements by CyTOF allow for a wide range of cells to be analyzed at once, due to the availability of a large staining panel without the complication of fluorescence spillover. These are also compatible with direct detection of integrated iodine signal, with minimal impact on immunophenotyping based on the surface markers. Mathematical modeling beyond a binary classification of surface marker abundance allows for a continuum of cellular states as the cells transition from one state to another. Thus, we present a complete and robust method for directly quantifying differentiation at the systemic level, allowing for system-wide comparisons between different mouse strains and/or experimental conditions. Published 2019. This article is a U.S. Government work and is in the public domain in the USA.

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Quantifying the Dynamics of Hematopoiesis by In Vivo IdU Pulse-Chase, Mass Cytometry, and Mathematical Modeling

Abstract

Bibliographical note

Keywords

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