Application Note: Magnetic Isolation and Rare Cell Gene Expression Analysis of Breast Cancer Stem Cells
11 Apr 2011
This application poster by Miltenyi Biotec addresses the challenges of cancer stem cell (CSC) isolation and amplification of small RNA amounts for transcriptome sequencing. A method is developed for the semi-automated dissociation of human tumor tissue yielding high recoveries of viable single cells for subsequent molecular analysis by high-throughput sequencing. Breast CSCs were isolated by depletion of CD24+ and CD45+ cells and subsequent enrichment of CD44+ cells. CSCs were enriched from 5% in the original fraction to a purity of 94%. mRNA from isolated breast CSCs was amplified by μMACS™ SuperAmp™ global PCR-based RNA amplification. Even from the small number of cells, the method allows for amplified cDNA to be analysed by high-throughput gene expression profiling.