OMIP-079
Cell cycle of CD4+ and CD8+ naïve/memory T cell subsets, and of Treg cells from mouse spleen
Cytometry PART A, Volume 99, Issue 12, 1171-1175 (2021)
Ambra Natalini,Sonia Simonetti,Gabriele Favaretto,Giovanna Peruzzi,Fabrizio Antonangeli,Angela Santoni,Miguel Muñoz-Ruiz,Adrian Hayday,Francesca Di Rosa
Description: A multicolor flow cytometry panel was designed and optimized to define the following nine mouse T cell subsets: Treg (CD3+ CD4+ CD8− FoxP3+), CD4+ T naïve (CD3+ CD4+ CD8−FoxP3− CD44int/low CD62L+), CD4+ T central memory (CD3+ CD4+ CD8− FoxP3− CD44high CD62L+), CD4+ T effector memory (CD3+ CD4+ CD8− FoxP3− CD44high CD62L−), CD4+ T EMRA (CD3+ CD4+ CD8− FoxP3− CD44int/low CD62L−), CD8+ T naïve (CD3+ CD8+ CD4− CD44int/low CD62L+), CD8+ T central memory (CD3+ CD8+ CD4− CD44high CD62L+), CD8+ T effector memory (CD3+ CD8+ CD4− CD44high CD62L−), and CD8+ T EMRA (CD3+ CD8+ CD4− CD44int/low CD62L−). In each T cell subset, a dual staining for Ki-67 expression and DNA content was employed to distinguish the following cell cycle phases: G0 (Ki67−, with 2n DNA), G1 (Ki67+, with 2n DNA), and S-G2/M (Ki67+, with 2n < DNA ≤ 4n). This panel was established for the analysis of mouse (C57BL/6J) spleen.
Species: Mouse
Tissue: Spleen
Cell Subset: T cell, Ctl, T helper, T memory, Treg
Keyword: Activation, Cell cycle, Proliferation
Machine: BD LSR Fortessa
Panel details
| Marker | Species | Clone | Fluorophore | Purpose | Co-expressions groups | Antigen Density |
|---|---|---|---|---|---|---|
| CD44 | Mouse | IM7 | APC | NaÏve/Memory subset identification | T cell | Med |
| FOXP3 | Mouse | FJK-16s | PE | Treg identification | T cell | Low |
| CD4 | Mouse | RM4-5 | PE-CF594 | Helper T cell identification | T cell | High |
| eFluor 780 Fix Viability | All Species | eFluor 780 Fix Viability | Live/Dead cell discrimination | |||
| CD3 | Mouse | 145-2c11 | FITC | Pan T cell marker | T cell | High |
| Ki-67 | Mouse | SolA15 | Alexa Fluor 700 | Quiescence/cell cycle | T cell | Low |
| Hoechst 33342 | All Species | Hoechst (33342) | DNA content/cell cycle | T cell | Unspecified | |
| CD62L | Mouse | Mel-14 | PE-Cy7 | NaÏve/Memory subset identification | T cell | Med |
| CD8a | Mouse | 53-6.7 | Brilliant Violet 785 | Cytotoxic T cell identification | T cell | High |
