A Technique to Generate a Human PBMC-Engrafted Humanized Xenograft Mouse Model

Published: May 31, 2024

Abstract

Source: Li, Z., et al. A Human Peripheral Blood Mononuclear Cell (PBMC) Engrafted Humanized Xenograft Model for Translational Immuno-oncology (I-O) Research. J. Vis. Exp. (2019)

This video demonstrates an assay to generate a humanized xenograft mouse model. An immunocompromised mouse is injected with cyclophosphamide to cause immune cell depletion. Patient-derived tumor cells mixed with human peripheral blood mononuclear cells (PBMCs) in a hydrogel are subcutaneously injected into the mouse, forming a tumor with infiltrating T cells.

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. Establishment of human PBMC-based model Myeloablation of NOD/SCID mice using cyclophosphamide: determination of optimal doses​ Purchase female nonobese diabetic/severe …

開示

The authors have nothing to disclose.

Materials

PBMC separation /cell culture
Histopaque-1077 Sigma 10771 Cell isolation
DMEM Corning 10-013-CVR Cell culture
DPBS Corning 21-031-CVR Cell culture
FBS Corning 35-076-CV Cell culture
Penicillin-Streptomycin, Liquid Gibco 15140-163 Cell culture
Trypsin-EDTA (0.25%), phenol red Gibco 25200-114 Cell culture
Matrigel Corning 356237 CDX inoculation
FACS analysis
Deoxyribonuclease I from bovine pancreas Sigma DN25 Sample preparation
Collagenase Type I Sigma C0130 Sample preparation
Anti-mouse/human CD11b (M1/70) antibody BioLegend 101206 FACS
Anti-mouse Ly-6C (HK1.4) antibody BioLegend 128008 FACS
Anti-mouse Ly-6G (1A8) antibody BioLegend 127614 FACS
Anti-human CD8 (OKT8) antibody Sungene Biotech H10082-11H FACS
Anti-human CD279 (MIH4) antibody eBioscience 12-9969-42 FACS
Anti-human CD3 (HIT3a) antibody 4A Biotech FACS
Guava easyCyte 8HT Benchtop Flow Cytometer Millipore 0500-4008 FACS
Tumor/PDX implantation /dosing / measurement
Cyclophosphamide J&K Cat#419656, CAS#6055-19-2 In vivo efficacy
Disulfiram J&K Cat#591123, CAS#97-77-8 In vivo efficacy
Syringe BD 300841 CDX inoculation
Hypodermic needles (14G) Shanghai SA Mediciall & Plastic Instruments Co., Ltd. 0.7*32 TW SB PDX inoculation
Vernier Caliper (MarCal) Mahr 16ER Tumor measurement
IVC individual ventilated cages Lingyunboji Ltd. IVC-128 Animal facility

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記事を引用
A Technique to Generate a Human PBMC-Engrafted Humanized Xenograft Mouse Model. J. Vis. Exp. (Pending Publication), e22231, doi: (2024).

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