Delivering Effector T Lymphocytes into Mice Carrying Human Brain Tumors

Published: May 31, 2024

Abstract

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.

Protocol

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 Prepare and irradiate feeder cells at 35 Gy. For the stimulation of 2 x 105 – 4 x 105 effector cells, prepare 10 x 10…

Representative Results

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.

Divulgaciones

The authors have nothing to disclose.

Materials

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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Delivering Effector T Lymphocytes into Mice Carrying Human Brain Tumors. J. Vis. Exp. (Pending Publication), e22239, doi: (2024).

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