Technical Note: R&D Systems Cell Selection Kits for Cancer-associated Cells: R&D Systems

MagCellect™ Human CD24^–CD44⁺ Breast Cancer Stem Cell Isolation Kit

R&D Systems offers several kits that are designed to isolate tumor associated cell populations, including a breast cancer stem cell isolation kit and multiple other kits that isolate cells expressing specific cancer-related markers. The MagCellect Human CD24^–CD44⁺ Breast Cancer Stem Cell Isolation Kit (Catalog # MAGH111) was designed to isolate a small population of human breast cancer cells that have the unique ability to form new tumors in mice.¹ This kit utilizes a two-step procedure that combines negative and positive selection techniques. CD24^low/– cells are initially enriched by negative selection whereby unwanted CD24⁺ cells are tagged and magnetically removed. CD44⁺ cells are subsequently isolated from the CD24^low/– population by positive selection using a biotinylated human CD44 antibody, streptavidin-conjugated magnetic beads, and a MagCellect Magnet (Catalog # MAG997, or equivalent). The efficiency of enrichment can be assessed by staining the recovered cells with fluorochrome-conjugated antihuman CD24 and CD44 detection antibodies provided in the kit.

Assay Principle

Analysis of CD24<sup>low/–</sup>CD44<sup>+</sup> Cells Isolated using the MagCellect Human CD24<sup>–</sup>CD44<sup>+</sup> Breast Cancer Stem Cell Isolation Kit.

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Analysis of CD24^low/–CD44⁺ Cells Isolated using the MagCellect Human CD24^–CD44⁺ Breast Cancer Stem Cell Isolation Kit. A small population of CD24^low/–CD44⁺ cells was isolated from MCF-7 human breast adenocarcinoma cells using the MagCellect CD24^–CD44⁺ Breast Cancer Stem Cell Isolation Kit (Catalog # MAGH111). CD24^low/–CD44⁺ cells (upper left quadrant), before (A) and after (B) enrichment, were detected by double-staining with APC-conjugated Mouse Anti-Human CD24 and PE-conjugated Mouse Anti-Human CD44 Detection Antibodies (both provided in the kit). A histogram profiling the enrichment of CD24^low/– cells (filled histogram) from the original cell population (open histogram) is shown (C).