Generating a Double Humanized BLT (Bone-Marrow, Liver, Thymus) Mouse Model

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

1. Purchase 6-8-week-old NSG mice (NOD.Cg-Prkdc^scidIl2rg^tm1Wjl/SzJ).
2. 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 pressure.
   1. House and maintain the mice in autoclaved individual microisolator cages in a rack system capable of managing air exchange with....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
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