TY - JOUR

T1 - Multiplex cell fate tracking by flow cytometry

AU - Rodríguez-Martínez, Marta

AU - Hills, Stephanie A.

AU - Diffley, John F.X.

AU - Svejstrup, Jesper Q.

N1 - Funding Information: This work was supported by the Francis Crick Institute (FCI), which receives its core funding from Cancer Research UK (FC001166), the UK Medical Research Council (FC001166), and the Wellcome Trust (FC001166), and by a grant from the European Research Council, Agreements 693327 (TRANSDAM) to JQS. We thank the flow cytometry facility of the Francis Crick Institute for their help and support, and especially Derek Davies for his ideas and input for this manuscript. Funding Information: Funding: This work was supported by the Francis Crick Institute (FCI), which receives its core funding from Cancer Research UK (FC001166), the UK Medical Research Council (FC001166), and the Wellcome Trust (FC001166), and by a grant from the European Research Council, Agreements 693327 (TRANSDAM) to JQS. Publisher Copyright: © 2020 by the authors. Licensee MDPI, Basel, Switzerland.

PY - 2020/9

Y1 - 2020/9

N2 - Measuring differences in cell cycle progression is often essential to understand cell behavior under different conditions, treatments and environmental changes. Cell synchronization is widely used for this purpose, but unfortunately, there are many cases where synchronization is not an option. Many cell lines, patient samples or primary cells cannot be synchronized, and most synchronization methods involve exposing the cells to stress, which makes the method incompatible with the study of stress responses such as DNA damage. The use of dual-pulse labelling using EdU and BrdU can potentially overcome these problems, but the need for individual sample processing may introduce a great variability in the results and their interpretation. Here, we describe a method to analyze cell proliferation and cell cycle progression by double staining with thymidine analogues in combination with fluorescent cell barcoding, which allows one to multiplex the study and reduces the variability due to individual sample staining, reducing also the cost of the experiment.

AB - Measuring differences in cell cycle progression is often essential to understand cell behavior under different conditions, treatments and environmental changes. Cell synchronization is widely used for this purpose, but unfortunately, there are many cases where synchronization is not an option. Many cell lines, patient samples or primary cells cannot be synchronized, and most synchronization methods involve exposing the cells to stress, which makes the method incompatible with the study of stress responses such as DNA damage. The use of dual-pulse labelling using EdU and BrdU can potentially overcome these problems, but the need for individual sample processing may introduce a great variability in the results and their interpretation. Here, we describe a method to analyze cell proliferation and cell cycle progression by double staining with thymidine analogues in combination with fluorescent cell barcoding, which allows one to multiplex the study and reduces the variability due to individual sample staining, reducing also the cost of the experiment.

KW - BrdU

KW - EdU

KW - Fluorescent cell barcoding

U2 - 10.3390/mps3030050

DO - 10.3390/mps3030050

M3 - Journal article

AN - SCOPUS:85090781500

VL - 3

SP - 1

EP - 9

JO - Methods and Protocols

JF - Methods and Protocols

SN - 2409-9279

IS - 3

M1 - 50

ER -