YILIMART - Mouse Cell Depletion Kit

The Mouse Cell Depletion Kit has been designed for the enrichment of untouched human cells upon xenotransplantation.

Background information

The Mouse Cell Depletion Kit has been designed for the enrichment of untouched human cells upon xenotransplantation. Human tumor xenografts represent the benchmark for research areas like drug discovery, cancer stem cell biology, and metastasis prediction

¹

. During the growth phase

in vivo

, xenografted tissue is vascularized and infiltrated by cells of mouse origin, including heterogeneous lymphocyte subpopulations, fibroblasts, and endothelial cells. The level of infiltration is highly dependent on multiple factors like tumor subtype, growth rate, and region of transplantation. However, even when these factors are kept constant, the amount and composition of infiltrating mouse cells are highly variable, which makes accurate molecular downstream analyses difficult. The contaminating mouse cells lead to cross-hybridization of mouse-derived molecules to human probes on microarrays and a significant reduction of sensitivity caused by measurement of mouse signals during next-generation sequencing or proteome analysis

²

. In addition, the culture of human tumor cells is frequently hampered by murine fibroblasts overgrowing the target cells. For optimal results, the Mouse Cell Depletion Kit should be used in combination with the Tumor Dissociation Kit, human (# 130-095-929).

Columns

LS or autoMACS

^®

Columns.

Figure 1

EpCAM positive human colon carcinoma cells were isolated from a heterogenous cell population using the Mouse Cell Depletion Kit, an LS Column, and a QuadroMACS™ Separator after the tissue was dissociated using the Tumor Dissociation Kit, human and gentleMACS Octo Dissociator. The figure shows the sample before separation and the isolated human tumor cells after the mouse cells have been depleted. The Labeling Check Reagent-APC was used to analyze mouse cells.

Unseparated fraction	Isolated human tumor cells

Figure 2

EpCAM positive human colon carcinoma cells were isolated from a heterogenous cell population and subsequently, the unseparated fraction, the human tumor cell fraction, and the mouse cell containing fraction were cultured for three days in expansion medium (90% RPMI-1640, 10% FBS, 100 U/mL penicillin, and 100 U/mL streptomycin). After fixation, cells were stained using antibodies specific for murine vimentin (red) and human EpCAM (green). Cell nuclei were stained with DAPI (blue).