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