Source: Jarry, U., et al. Stereotactic Adoptive Transfer of Cytotoxic Immune Cells in Murine Models of Orthotopic Human Glioblastoma Multiforme Xenografts. J. Vis. Exp. (2018).
This video demonstrates a stereotaxic injection technique for administering human effector T lymphocytes into a murine model carrying a human brain tumor. Upon positioning a tumor-bearing anesthetized mouse on a stereotactic frame and creating a burr hole at the injection location on the skull, effector T lymphocytes are injected at the tumor site.
All procedures involving sample collection have been performed in accordance with the institute's IRB guidelines. All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Non-specific Expansion of Cytotoxic Effector T Lymphocytes
2. Pre-operative Effector Cells Preparation
3. Stereotactic Injection
Figure 1: Picture of the main anatomical landmarks on the mouse skull. This includes sagittal (red line) and coronal (blue line) sutures and their intersection (Bregma) used to orient the site of injection. The scale bar is indicated.
The authors have nothing to disclose.
PBMCs | from 3 different healthy donors | ||
BLCLs | from 3 different donors | ||
Roswell Park Memorial Institute medium (RPMI) | Gibco | 31870-025 | |
FCS | Dutscher | S1810-500 | |
L-glutamine | Gibco | 25030-024 | |
penicillin/streptomycin | Gibco | 15140-122 | |
IL-2 | Novartis | proleukin | |
PHA-L | Sigma | L4144 | |
Stereotaxic frame | Stoelting Co. | 51600 | |
Mouse adaptator for stereotaxic frame | Stoelting Co. | 51624 | |
microsyringe pump injector | WPI | UMP3-4 | |
NanoFil Syringe | WPI | NF34BV-2 | |
NSG mice | Charles River | NSGSSFE07S | |
Ketamine | Merial | Imalgène 1000 | |
Xylazine | Bayer | Rompur 2% | |
Scissors | WPI | 201758 | |
Forceps | WPI | 501215 | |
OmniDrill 115/230V | WPI | 503598 | |
Vicryl 4-0 | Ethicon | VCP397H | |
Xylocaine | Astrazeneca | 3634461 |
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