Isolating Cord Blood Hematopoietic Stem Cells to Generate Human Immune System Mice

Published: April 30, 2024

Abstract

Source: Lanis, J. M., et al. Testing Cancer Immunotherapeutics in a Humanized Mouse Model Bearing Human Tumors. J. Vis. Exp. (2022)

This video demonstrates a technique for generating human immune system (HIS) mice. CD34+ hematopoietic stem cells (HSCs) are isolated from human cord blood buffy coat cells using antibody-coated magnetic beads. The CD34+ HSCs are expanded in a growth medium enriched with cytokines and cryopreserved. Finally, the thawed HSCs are injected into an immunodeficient mouse pup to establish the HIS mouse model.

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. Generation of HIS mice Mouse husbandry of BALB/c -Rag2null Il2RγCnull SirpαNOD (BRGS) mice NOTE: This strain is extremely immun…

Offenlegungen

The authors have nothing to disclose.

Materials

1 mL syringe w/needles McKesson 1031815
15 mL tubes Grenier Bio-One 188271
2-mercaptoethanol Sigma M6250
50 mL tubes Grenier Bio-One 227261
AutoMACS Pro Separator Miltenyi 130-092-545
BSA Fisher Scientific BP1600100
Chill 15 Rack Miltenyi 130-092-952
Embryonic Stemcell FCS Gibco 10439001
Eppendorf Tubes; 1.5 mL volume Grenier Bio-One 616201
Excel Microsoft
FBS Benchmark 100-106 500mL
Ficoll Hypaque GE Healthcare 45001752
glass pipettes DWK Life Sciences 63A53
HBSS w/ Ca & Mg Sigma 55037C
MultiRad 350 Precision X-Ray
PBS Corning 45000-446
PRISM Graphpad
Rec Hu FLT3L R&D systems 308-FK-005/CF
Rec Hu IL6 R&D systems 206-IL-010/CF
Rec Hu SCF R&D systems 255SC010
Serological pipettes 25 mL Fisher Scientific 1367811
Sterile filter Nalgene 567-0020
Sterile molecular water Sigma 7732-18-5
Yeti Cell Analyzer Bio-Rad 12004279

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Isolating Cord Blood Hematopoietic Stem Cells to Generate Human Immune System Mice. J. Vis. Exp. (Pending Publication), e22227, doi: (2024).

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