Measurement of Mitochondrial Mass and Membrane Potential in Hematopoietic Stem Cells and T-cells by Flow Cytometry.

Details

Serval ID
serval:BIB_DE9A27B5B4E4
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Measurement of Mitochondrial Mass and Membrane Potential in Hematopoietic Stem Cells and T-cells by Flow Cytometry.
Journal
Journal of visualized experiments
Author(s)
Girotra M., Thierry A.C., Harari A., Coukos G., Naveiras O., Vannini N.
ISSN
1940-087X (Electronic)
ISSN-L
1940-087X
Publication state
Published
Issued date
26/12/2019
Peer-reviewed
Oui
Number
154
Language
english
Notes
Publication types: Journal Article ; Video-Audio Media
Publication Status: epublish
Abstract
A fine balance of quiescence, self-renewal, and differentiation is key to preserve the hematopoietic stem cell (HSC) pool and maintain lifelong production of all mature blood cells. In recent years cellular metabolism has emerged as a crucial regulator of HSC function and fate. We have previously demonstrated that modulation of mitochondrial metabolism influences HSC fate. Specifically, by chemically uncoupling the electron transport chain we were able to maintain HSC function in culture conditions that normally induce rapid differentiation. However, limiting HSC numbers often precludes the use of standard assays to measure HSC metabolism and therefore predict their function. Here, we report a simple flow cytometry assay that allows reliable measurement of mitochondrial membrane potential and mitochondrial mass in scarce cells such as HSCs. We discuss the isolation of HSCs from mouse bone marrow and measurement of mitochondrial mass and membrane potential post ex vivo culture. As an example, we show the modulation of these parameters in HSCs via treatment with a metabolic modulator. Moreover, we extend the application of this methodology on human peripheral blood-derived T cells and human tumor infiltrating lymphocytes (TILs), showing dramatic differences in their mitochondrial profiles, possibly reflecting different T cell functionality. We believe this assay can be employed in screenings to identify modulators of mitochondrial metabolism in various cell types in different contexts.
Pubmed
Web of science
Create date
17/01/2020 17:30
Last modification date
15/07/2020 6:26
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