Fluorescence-Activated Cell Sorting of Murine Mammary Cancer Stem-Like Cell Subpopulations with HIF Activity

摘要: Fluorescence-activated cell sorting (FACS) is a common method to identify and isolate subpopulations within complex mixture of cells based on their light scatter fluorescent staining profiles. FACS widely used enrich for normal tissue tumor that have stem potential. Whereas protocols using conventional breast cancer lines are relatively routine, additional technical challenges encountered when populations from freshly digested solid tumors, particularly use in downstream (CSC) assays. First, it more difficult live, single whole second, prepared enzymatically tumors typically sensitive death following the physical stresses digestion, pipetting, sorting. Herein methods described been optimized harvest profile viable epithelial late-stage mammary originating mouse virus (MMTV)-polyomavirus middle T antigen (PyMT) transgenic mouse. Protocols were designed single, viable, MMTV-PyMT sorted by facilitate collection suitable head-to-head comparison CSC activity tumorsphere assays vitro or limiting dilution transplantation vivo.