Modified cell trace violet proliferation assay preserves lymphocyte viability and allows spectral flow cytometry analysis
Details
Publication Year 2024-05,Volume 105,Issue #5,Page 394-403
Journal Title
Cytometry Part A
Publication Type
Research article
Abstract
In this study we describe three different methods for labeling T lymphocytes with cell trace violet (CTV), in order to track cell division in mouse and human cells, in both the in vitro and in vivo setting. We identified a modified method of CTV labeling that can be applied directly to either conventional or spectral flow cytometry, that maintained lymphocyte viability and function, yet minimized dye spill-over into other fluorochrome channels. Our optimized method for CTV labeling allowed us to identify up to eight cell divisions and the replication index for in vitro-stimulated mouse and human lymphocytes, and the co-expression of T-cell subset markers. Furthermore, the homeostatic trafficking, expansion and division of CTV-labeled congenic donor T cells could be detected using spectral cytometry, in an adoptive T-cell transfer mouse model. Our optimized CTV method can be applied to both in vitro and in vivo settings to examine the behavior and phenotype of activated T cells.
Publisher
Wiley
Keywords
Animals; *Flow Cytometry/methods; Humans; Mice; *Cell Survival; *Cell Proliferation; Staining and Labeling/methods; Mice, Inbred C57BL; Lymphocyte Activation/immunology; T-Lymphocytes/cytology/immunology; Fluorescent Dyes/chemistry; cell trace violet; congenic adoptive transfer mouse model; conventional flow cytometry; lymphocyte proliferation; spectral flow cytometry
Department(s)
Clinical Haematology
Open Access at Publisher's Site
https://doi.org/10.1002/cyto.a.24830
Terms of Use/Rights Notice
Refer to copyright notice on published article.


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Last Modified: 2024-07-04 04:51:28

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