Isolating Invariant Natural Killer T Cells from a Mouse Model

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

1. Directional induction of iNKT cells
   1. Inject normal mice intraperitoneally with α-Galactosylceramide (α-GalCer) (0.1 mg/kg of body weight).
2. Isolation of iNKT cells
   1. Three days after modeling, inject mice intraperitoneally with 1% sodium pentobarbital (50 mg/kg of body weight) for anesthetizatio....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
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