Delivering Effector T Lymphocytes into Mice Carrying Human Brain Tumors

In This Article

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 10⁶ peripheral-blood mononuclear cells (PBMCs) and 1 x 10⁶ B-lymphoblastoid cell lines (BLCLs) from three distinct, healthy donors.

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Results

Stereotaxic surgery on a mouse brain, surgical methodology, neuroscience research experiment.
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.

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