Abstract
CellTrace™ Violet (CTV) is a powerful tool for tracking cell proliferation by permanently binding cellular proteins and rendering the cell fluorescent. After cell division, each daughter cell contains half of the parent cell’s fluorescence, enabling quantification of proliferation via flow cytometry. This method enables monitoring of several generations of cell division and tracking of different cell populations in co-culture. Here we describe the use of CellTrace™ Violet in different cell types, and we share important observations we made during protocol optimization.
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Notes
- 1.
Compatibility criteria will be discussed in Subheading 3.2.3.
References
Begum J, Day W, Henderson C, Purewal S, Cerveira J, Summers H, Rees P, Davies D, Filby A (2013) A method for evaluating the use of fluorescent dyes to track proliferation in cell lines by dye dilution. Cytometry A 83(12):1085–1095. https://doi.org/10.1002/cyto.a.22403
Quah BJ, Parish CR (2012) New and improved methods for measuring lymphocyte proliferation in vitro and in vivo using CFSE-like fluorescent dyes. J Immunol Methods 379(1–2):1–14. https://doi.org/10.1016/j.jim.2012.02.012
Tempany JC, Zhou JH, Hodgkin PD, Bryant VL (2018) Superior properties of CellTrace Yellow as a division tracking dye for human and murine lymphocytes. Immunol Cell Biol 96(2):149–159. https://doi.org/10.1111/imcb.1020
Winter S, Martin E, Boutboul D, Lenoir C, Boudjemaa S, Petit A, Picard C, Fischer A, Leverger G, Latour S (2018) Loss of RASGRP1 in humans impairs T-cell expansion leading to Epstein-Barr virus susceptibility. EMBO Mol Med 10(2):188–199. https://doi.org/10.15252/emmm.201708292
Perego M, Maurer M, Wang JX, Shaffer S, Muller AC, Parapatics K, Li L, Hristova D, Shin S, Keeney F, Liu S, Xu X, Raj A, Jensen JK, Bennett KL, Wagner SN, Somasundaram R, Herlyn M (2018) A slow-cycling subpopulation of melanoma cells with highly invasive properties. Oncogene 37(3):302–312. https://doi.org/10.1038/onc.2017.341
Zhou P, Chen X, Li M, Sun X, Tan J, Wang X, Chu Y, Zhang Y, Cheng T, Zhou J, Wang G, Yuan W (2019) Overexpression of PRDM5 promotes acute myeloid leukemia cell proliferation and migration by activating the JNK pathway. Cancer Med 8(8):3905–3917. https://doi.org/10.1002/cam4.2261
Filby A, Begum J, Jalal M, Day W (2015) Appraising the suitability of succinimidyl and lipophilic fluorescent dyes to track proliferation in non-quiescent cells by dye dilution. Methods 82:29–37. https://doi.org/10.1016/j.ymeth.2015.02.016
Filby A, Day W, Purewal S, Martinez-Martin N (2016) The analysis of cell cycle, proliferation, and asymmetric cell division by imaging flow cytometry. In: Bartneva NS, Vorobjev IA (eds) Imaging flow cytometry methods and protocols. Methods in molecular biology, vol 1389, 1st edn. Humana Press, New York, pp 71–95. https://doi.org/10.1007/978-1-4939-3302-0
Lacy KJ, Parlane NA, Riley CB, Gee EK, Roberts JM, McIlwraith CW (2020) CellTrace Violet inhibits equine lymphocyte proliferation. Vet Immunol Immunopathol 223:110037. https://doi.org/10.1016/j.vetimm.2020.110037
CellTrace™ Cell Proliferation Kits user guide (2017) vol MAN0002595, E.00 edn. ThermoFisher Scientific
Tario JD Jr, Conway AN, Muirhead KA, Wallace PK (2018) Monitoring cell proliferation by dye dilution: considerations for probe selection. Methods Mol Biol 1678:249–299. https://doi.org/10.1007/978-1-4939-7346-0_12
McKinnon KM (2018) Flow cytometry: an overview. Curr Protoc Immunol 120:5 1 1–5 1 11. https://doi.org/10.1002/cpim.40
Haramoto Y, Onuma Y, Mawaribuchi S, Nakajima Y, Aiki Y, Higuchi K, Shimizu M, Tateno H, Hirabayashi J, Ito Y (2020) A technique for removing tumourigenic pluripotent stem cells using rBC2LCN lectin. Regen Ther 14:306–314. https://doi.org/10.1016/j.reth.2020.03.017
Ivanov DP, Parker TL, Walker DA, Alexander C, Ashford MB, Gellert PR, Garnett MC (2015) In vitro co-culture model of medulloblastoma and human neural stem cells for drug delivery assessment. J Biotechnol 205:3–13. https://doi.org/10.1016/j.jbiotec.2015.01.002
Chung S, Kim SH, Seo Y, Kim SK, Lee JY (2017) Quantitative analysis of cell proliferation by a dye dilution assay: application to cell lines and cocultures. Cytometry A 91(7):704–712. https://doi.org/10.1002/cyto.a.23105
Siegers GM, Ribot EJ, Keating A, Foster PJ (2013) Extensive expansion of primary human gamma delta T cells generates cytotoxic effector memory cells that can be labeled with Feraheme for cellular MRI. Cancer Immunol Immunother 62(3):571–583. https://doi.org/10.1007/s00262-012-1353-y
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Lemieszek, M.B., Findlay, S.D., Siegers, G.M. (2022). CellTrace™ Violet Flow Cytometric Assay to Assess Cell Proliferation. In: Christian, S.L. (eds) Cancer Cell Biology. Methods in Molecular Biology, vol 2508. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2376-3_9
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DOI: https://doi.org/10.1007/978-1-0716-2376-3_9
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