Protocol

Basic Cell Culture Protocols

Volume 946 of the series Methods in Molecular Biology pp 395-409

Date:

Enzymatic Dissociation, Flow Cytometric Analysis, and Culture of Normal Mouse Mammary Tissue

  • Michael PraterAffiliated withCancer Research, UK Cambridge Research Institute
  • , Mona ShehataAffiliated withCancer Research UK, Cambridge Research Institute
  • , Christine J. WatsonAffiliated withDepartment of Pathology, University of Cambridge
  • , John StinglAffiliated withCancer Research UK, Cambridge Research Institute Email author 

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Abstract

Evidence is emerging that the mouse mammary epithelium is arranged as a hierarchy that spans from stem cells to lineage-restricted progenitor cells to differentiated luminal and myoepithelial cells. The use of fluorescence-activated cell sorting (FACS) in combination with quantitative functional clonal assays represents a powerful tool for studying the properties of mouse mammary stem and progenitor cells. This chapter outlines the experimental procedures for generating single viable cell suspensions of mouse mammary epithelial cells, immunostaining cells for flow cytometry, in vitro assays for the detection and enumeration of mouse mammary progenitor cells, and in vivo assays for the detection and enumeration of mouse mammary stem cells.

Key words

Mouse mammary gland Stem cells Flow cytometry Cell culture