Isolating Cord Blood Hematopoietic Stem Cells to Generate Human Immune System Mice
Transcript
Begin with human umbilical cord blood cells, including CD34+ hematopoietic stem cells, or HSCs.
Add antibodies to block cellular Fc receptors, preventing non-specific binding of HSC-specific magnetic beads.
Introduce the antibody-coated magnetic beads targeting CD34+ HSCs. Centrifuge and discard the supernatant.
Resuspend the cells in a buffer. Pass them through a magnetic separator to retain bead-bound HSCs; unbound cells flow through.
Remove the magnetic field to collect the HSCs.
Centrifuge and resuspend the cells in a growth medium. Plate the cells and incubate.
The medium contains growth factors and cytokines that promote HSC survival and expansion.
Harvest the cells. Add a freezing medium, transfer to a cryovial, and freeze.
To generate a human immune system, or HIS, mouse model, thaw the HSCs. Add them to a pre-warmed medium.
Centrifuge and resuspend the cells in a buffer.
The cells are ready for injection into an immunodeficient mouse pup for HIS model generation.
