Generating a Double Humanized BLT (Bone-Marrow, Liver, Thymus) Mouse Model
Published: May 31, 2024
Abstract
Source: Daharsh, L. et al., A Double Humanized BLT-mice Model Featuring a Stable Human-Like Gut Microbiome and Human Immune System. J. Vis. Exp. (2019)
This video demonstrates the method for generating a double humanized bone BLT (marrow, liver, and thymus) mouse with a functional human immune system and a stable engrafted human-like gut microbiome.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Mice housing and maintenance Purchase 6-8-week-old NSG mice (NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ). House the mice under SPF conditions with air exchange, prefilters, and high-efficiency particulate air (HEPA) filters (0.22 μm) in a room with controlled temperature, humidity, and…
Disclosures
The authors have nothing to disclose.
Materials
| Animal Feeding Needles 18G | Cadence Science | 9928B | |
| Clidox-s Activator | Pharmacal Research Laboratories | 95120F | |
| Clidox-s Base | Pharmacal Research Laboratories | 96125F | |
| DGM 108 cage rack | Techniplast | ||
| Flat Brown Grocery Bag 3-5/8"D x 6"W x 11-1/16"L | Grainger | 12R063 | |
| FMT Upper Delivery Microbiota Preparations | OpenBiome | FMP30 | |
| Grape Kool-Aid | Kraft Foods Inc. | ||
| hCD19-PE/Cy5 | Biolegend | 302209 | |
| hCD3-PE | Biolegend | 300408 | |
| hCD4-Alexa 700 | Biolegend | 300526 | |
| hCD45-FITC | Biolegend | 304006 | |
| hCD8-APC/Cy7 | Biolegend | 301016 | |
| Lactate Buffered Ringer's Solution | Boston BioProducts Inc | PY-906-500 | |
| mCD45-APC | Biolegend | 103111 | |
| Microvette 100 K3E | Microvette | 20.1278.100 | |
| Neosporin First Aid Antibiotic/Pain Relieving Ointment | Neosporin | ||
| NSG mice (NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ) | The Jackson Laboratory | 5557 | |
| PrecisionGlide 25 G Needle | BD | 305127 | |
| RS200 X-ray irradiator | RAD Source Technologies | ||
| Sealsafe Plus GM500 microisolator cages | Techniplast | ||
| Sterile Non-woven Gauze | Fisherbrand | 22-028-558 | |
| Teklad global 16% protein irradiated mouse chow | Teklad | 2916 |
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Cite This Article
Generating a Double Humanized BLT (Bone-Marrow, Liver, Thymus) Mouse Model. J. Vis. Exp. (Pending Publication), e22243, doi: (2024).