A Technique to Generate a Human PBMC-Engrafted Humanized Xenograft Mouse Model

Take an immunocompromised mouse possessing only neutrophils and monocytes as functional immune cells.

Intraperitoneally inject cyclophosphamide, a drug that targets immune progenitor cells.

The drug enters actively proliferating immune progenitor cells and crosslinks nucleic acids, causing cell death. The death of progenitor cells leads to immune cell depletion.

Orally administer disulfiram, a drug that forms an adduct with cyclophosphamide, reducing cyclophosphamide-induced side effects.

Subcutaneously inject a mixture of human tumor cells and peripheral blood mononuclear cells, or PBMCs, in a suitable gelatinous protein mixture.

The protein mixture forms a gel, mimicking the physiological extracellular matrix, and entraps the tumor cells.

Functional immune cell depletion allows the tumor cells to proliferate and form new blood vessels, transforming into a solid tumor xenograft.

Simultaneously, T cells from the injected PBMCs infiltrate the tumor.

The tumor xenograft exhibits growth and tumor-infiltrating T cells, establishing a humanized xenograft mouse model.

For myeloid cell depletion, intraperitoneally deliver 100 milligrams per kilogram of cyclophosphamide, and orally deliver 125 milligrams per kilogram of disulfiram to 6 to 8-week-old NOD/SCID female mice once a day for two days. 24 hours after the second dose of cyclophosphamide and disulfiram, resuspend 5 x 10⁶ freshly isolated human PBMC and 2.5 x 10⁶ human tumor cell line cells in 200 microliters of PBS plus 50% matrigel per animal, and inject the entire volume subcutaneously into the right flank of each experimental animal. Measure and record the primary tumor volume 2 times a week for 4 to 6 weeks.