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  • Enzymatic dissociation, flow cytometric analysis, and culture of normal mouse mammary tissue.

Enzymatic dissociation, flow cytometric analysis, and culture of normal mouse mammary tissue.

Methods in molecular biology (Clifton, N.J.) (2012-11-28)
Michael Prater, Mona Shehata, Christine J Watson, John Stingl
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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Hyaluronidase from bovine testes, Type IV-S, lyophilized powder (essentially salt-free), 750-3000 units/mg solid
Sigma-Aldrich
Hyaluronidase from sheep testes, Type II, lyophilized powder, ≥300 units/mg solid
Sigma-Aldrich
Hyaluronidase from sheep testes, Type V, lyophilized powder, ≥1,500 units/mg solid
Sigma-Aldrich
Hyaluronidase from bovine testes, Type VI-S, lyophilized powder, 3,000-15,000 units/mg solid
Sigma-Aldrich
Hyaluronidase from bovine testes, Type IV-S, powder, suitable for mouse embryo cell culture, 750-3000 units/mg solid
Sigma-Aldrich
Hyaluronidase from bovine testes, Type VIII, lyophilized powder, 300-1,000 U/mg
Sigma-Aldrich
Hyaluronidase from bovine testes, Type I-S, lyophilized powder, 400-1000 units/mg solid