Isolating Invariant Natural Killer T Cells from a Mouse Model

Published: April 30, 2024

Abstract

Source: Meng, M., et al. Adoptive Immunotherapy of iNKT Cells in Glucose-6-Phosphate Isomerase (G6PI)-Induced RA Mice. J. Vis. Exp. (2020)

This video illustrates a method for isolating invariant natural killer T cells, also known as iNKT cells. The procedure begins with injecting a synthetic glycolipid into a mouse to stimulate the activation and proliferation of iNKT cells in the spleen. Subsequently, single-cell suspensions are prepared from the spleen and incubated with iNKT-cell-recognizing complexes labeled with fluorophores, followed by magnetic bead-based separation, to isolate the iNKT cells.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Obtaining invariant natural killer T cells (iNKT Cells) with Adoptive Cellular Therapy Directional induction of iNKT cells Inject normal mice intraperitoneally with α-Galactosylceramide (α-GalCer) (0.1 mg/kg of body weight). Isolation of iNKT cells</…

Declarações

The authors have nothing to disclose.

Materials

Anti-PE MicroBeads Miltenyi 130-048-801 Germany
Columns Miltenyi MS Germany
Cryogenic Centrifuge Beckman Allegra® X-15R America
Mouse CD1d Tetramer-PE MBL TS-MCD-1 Japan
Mouse percoll Solarbio P8620 China
Optical Microscope Olympus Olympus-II Japan

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Isolating Invariant Natural Killer T Cells from a Mouse Model. J. Vis. Exp. (Pending Publication), e22221, doi: (2024).

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