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 |
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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 | |
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