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</…
開示
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 |
