Humanized Mice

Human cord blood was obtained from the St. Louis Cord Blood Bank. CD34⁺ cells were enriched using Dynabeads CD34 Positive Isolation Kit (Life Technologies) per the manufacturer’s instructions. 1 × 10⁵ cells were injected retro-orbitally into unconditioned NSG-W41 recipients. At least 2mo post-transplantation, mice were bled to confirm human chimerism. Peripheral blood was collected in 10mM EDTA/PBS via tail venipuncture of warmed mice. Red blood cells were lysed with 0.15M NH₄Cl, 10mM KHCO₃, 0.1mM EDTA, pH 7.2 solution (ACK) prior to antibody staining. For splenic chimerism, spleens were harvested and dissociated using frosted glass microscope slides. Red blood cells were lysed using ACK. Cells were filtered through 70μm nylon mesh prior to antibody staining.