Delivering Effector T Lymphocytes into Mice Carrying Human Brain Tumors

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.

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