Culturing of Activated T Cells from Human Peripheral Blood Mononuclear Cells
Published: July 31, 2023
Abstract
Source: Mølgaard, K., et al. Real-time Monitoring of Mitochondrial Respiration in Cytokine-differentiated Human Primary T Cells. J. Vis. Exp. (2021).
In this video, we describe a protocol for human primary T cell activation. The incubation of naïve T cells with magnetic beads coated with anti-CD3 and anti-CD28 antibodies, followed by interleukin-2 or interleukin-15 treatment, leads to the activation of naïve T cells and their differentiation and proliferation into effector or memory T cells.
Protocol
All procedures involving human participants have been performed in compliance with the institutional, national, and international guidelines for human welfare and have been reviewed by the local institutional review board. 1. Culturing of activated human primary T lymphocytes Thawing of cells (Day 1) Pre-warm 10 mL of RPMI 1640 per sample to around 37 °C. Take the desired number of peripheral blood mononuclear cell ampules and store them…
開示
The authors have nothing to disclose.
Materials
| 24-well tissue culture plate | Nunc | 142485 | |
| Anti-CD3xCD28 beads | Gibco | 11161D | |
| X-VIVO 15 | Lonza | BE02-060F | |
| Human Serum | Sigma Aldrich | H4522 | Heat inactivated at 56 °C for 30 min |
| IL-15 | Peprotech | 200-02 | |
| IL-2 | Peprotech | 200-15 | |
| Lymphoprep | Stemcell Technologies | 7801 | |
| PBS | Thermo Fisher | 10010023 | |
| RPMI 1640 | Gibco-Thermo Fisher | 61870036 | |
| T cell beads magnet DynaMag-2 Magnet | Thermo Fisher | 12321D |
