Home › Immunology › Depletion of Mouse Cells from Human Tumor Xenografts Significantly Improves Downstream Analysis of Target Cells
Immunology JoVE (Open Access) Citable · DOI

Depletion of Mouse Cells from Human Tumor Xenografts Significantly Improves Downstream Analysis of Target Cells

DOI: 10.3791/54259-v
What you'll learn
  • ✓ Perform enzymatic and mechanical dissociation of human tumor xenografts
  • ✓ Apply magnetic cell sorting to deplete mouse stromal and immune cells
  • ✓ Obtain pure human tumor cell populations for downstream analysis
Protocol

Human tumor xenografts are vascularized and infiltrated by cells of mouse origin during the growth phase in vivo. To circumvent the bias caused by these contaminating cells during downstream analysis, we developed a method allowing for comprehensive depletion of all mouse cells by magnetic cell sorting.

Difficulty
intermediate
Total time
~3–4 hours per tumor xenograft
Model organism
Mouse (human tumor xenograft model)
Biosafety
BSL-1

Steps

1
Dissociate tumor xenograft into single-cell suspension

Enzymatically and mechanically dissociate excised human tumor xenograft tissue to generate a single-cell suspension containing both human and mouse-derived cells.

▶ 00:40
2
Deplete mouse cells via magnetic cell separation

Use magnetic cell sorting technology to selectively remove all mouse-origin cells (stromal, immune, endothelial) from the dissociated tumor cell suspension, yielding enriched human tumor cells.

▶ 02:06
3
Verify mouse-cell depletion and analyze human cells

Perform flow cytometry, RNA-seq, or other downstream analyses on the mouse-depleted fraction to confirm removal of contaminating cells and characterize purified human tumor cell populations.

▶ 04:01
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