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

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記事を引用
Culturing of Activated T Cells from Human Peripheral Blood Mononuclear Cells. J. Vis. Exp. (Pending Publication), e21529, doi: (2023).

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