Constructing a Humanized Mouse Model with Engineered Immunity against HIV

Published: May 31, 2024

Abstract

Source: Zhen, A., et al. Stem-cell Based Engineered Immunity Against HIV Infection in the Humanized Mouse Model. J. Vis. Exp. (2016).

The video demonstrates a technique to generate a humanized mouse model with stem-cell-based engineered immunity against the human immunodeficiency virus (HIV). Human fetal thymus tissue and engineered hematopoietic stem cells (HSCs) are implanted into the kidney capsule of a severely immunocompromised mouse. The HSCs, which express chimeric antigen receptors (CARs) against HIV, are mixed in a gelatinous protein mixture that promotes their co-localization with thymus tissue. Post-procedure, the HSCs differentiate into various human immune cells, resulting in the development of a humanized mouse model with a functional human immune system against HIV.

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. Construction of Humanized Mice Engineered with CD4 Chimeric Antigen Receptor Processing Fetal Thymus and Isolating CD34+ HSCs from Fetal Liver Thymus Process…

Offenlegungen

The authors have nothing to disclose.

Materials

CD34 microbead kit miltenyi 130-046-702 For sorting human CD34+ progenitor cells
Bambanker Wako 302-14681 for freezing cells
QIAamp Viral RNA kit Qiagen 52904 For measuring viral load in the serum
MACSQuant Flow Cytometer Miltenyi For flow analysis
BD LSRFortessa™ BD biosciences For flow analysis
Hyaluronidase Sigma H6254-500MG For tissue digestion
Deoxyribonuclease I Worthington LS002006 for tissue digestion
Collagenase Life technology 17104-019 for tissue digestion
CFX Real time PCR detection system Biorad For measuring viral load and gene expression
Mice, strain NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ The Jackson Laboratory 5557 For constructing the humanized mice
Penicillin Streptomycin (Pen Strep) Thermo Fisher Scientific 10378016 For culturing cells
piperacillin/tazobactam Pfizer Zosyn Anti-fungal
Amphotericin B (Fungizone antimycotic) Thermo Fisher Scientific 15290-018 Anti-fungal
AUTOCLIP Wound Clips, 9 mm – 1000 units Becton Dickinson 427631 For surgery
Sterile Poly-Reinforced Aurora Surgical Gowns, 30 per case Medline DYNJP2707 For surgery
Sutures, 4-0, vicryl Owens and Minor 23000J304H For surgery
Alcohol prep pads Owens and Minor 3583006818 For surgery
Gloves, surgical, 6 1/2 Owens and Minor 4075711102 For surgery
Yssel's Serum-Free T-Cell Medium Gemini Bio-products 400-102 For CD34+ cell transduction
Human Serum Albumin Sigma-Aldrich A9511 For CD34+ cell transduction

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Diesen Artikel zitieren
Constructing a Humanized Mouse Model with Engineered Immunity against HIV. J. Vis. Exp. (Pending Publication), e22269, doi: (2024).

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