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…
Divulgazioni
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