3D culture assays of murine mammary branching morphogenesis and epithelial invasion

Kim Vy Nguyen-Ngoc, Eliah R. Shamir, Robert J. Huebner, Jennifer N. Beck, Kevin J. Cheung, Andrew Ewald

Research output: Contribution to journalArticle

Abstract

Epithelia are fundamental tissues that line cavities, glands, and outer body surfaces. We use three-dimensional (3D) embedded culture of primary murine mammary epithelial ducts, called “organoids,” to recapitulate in days in culture epithelial programs that occur over weeks deep within the body. Modulating the composition of the extracellular matrix (ECM) allows us to model cell- and tissue-level behaviors observed in normal development, such as branching morphogenesis, and in cancer, such as invasion and dissemination. Here, we describe a collection of protocols for 3D culture of mammary organoids in different ECMs and for immunofluorescence staining of 3D culture samples and mammary gland tissue sections. We illustrate expected phenotypic outcomes of each assay and provide troubleshooting tips for commonly encountered technical problems.

Original languageEnglish (US)
Pages (from-to)135-162
Number of pages28
JournalMethods in molecular biology (Clifton, N.J.)
Volume1189
DOIs
StatePublished - 2015

Fingerprint

Morphogenesis
Organoids
Breast
Human Mammary Glands
Fluorescent Antibody Technique
Extracellular Matrix
Epithelium
Staining and Labeling
Neoplasms

Keywords

  • 3D culture
  • Branching morphogenesis
  • Collagen I
  • Extracellular matrix
  • Immunofluorescence
  • Mammary epithelium
  • Matrigel
  • Organoids
  • Organotypic culture

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Medicine(all)

Cite this

3D culture assays of murine mammary branching morphogenesis and epithelial invasion. / Nguyen-Ngoc, Kim Vy; Shamir, Eliah R.; Huebner, Robert J.; Beck, Jennifer N.; Cheung, Kevin J.; Ewald, Andrew.

In: Methods in molecular biology (Clifton, N.J.), Vol. 1189, 2015, p. 135-162.

Research output: Contribution to journalArticle

Nguyen-Ngoc, Kim Vy ; Shamir, Eliah R. ; Huebner, Robert J. ; Beck, Jennifer N. ; Cheung, Kevin J. ; Ewald, Andrew. / 3D culture assays of murine mammary branching morphogenesis and epithelial invasion. In: Methods in molecular biology (Clifton, N.J.). 2015 ; Vol. 1189. pp. 135-162.
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