Journal of Mammary Gland Biology and Neoplasia

, 14:381

Adipose-Depleted Mammary Epithelial Cells and Organoids

  • Michael C. Rudolph
  • Elizabeth A. Wellberg
  • Steven M. Anderson
Article

DOI: 10.1007/s10911-009-9161-5

Cite this article as:
Rudolph, M.C., Wellberg, E.A. & Anderson, S.M. J Mammary Gland Biol Neoplasia (2009) 14: 381. doi:10.1007/s10911-009-9161-5

Abstract

Analysis of genes and proteins involved in lipid biosynthesis in mammary epithelial cells (MECs) is complicated by the presence of adipose tissue in the mammary gland, which may be predominant in whole tissue lysates depending upon developmental stage. We have developed a method based on protocols used to establish primary mammary epithelial cell cultures that allows for analysis of MECs depleted of adipose. Adipose depletion yields enriched MECs that are suitable for gene expression profiling and protein analysis from a single mouse. Additionally, the phosphorylation of proteins is maintained, allowing investigation of signal transduction events. Application of this method to the analysis of MECs from genetically modified mice will aid in the identification of factors controlling tissue-specific events in the mammary gland. In contrast to other methods such as laser capture microdissection, the MEC enrichment method described here is performed using standard lab supplies, equipment, and techniques.

Keywords

Mammary epithelial cellEnrichmentPregnancyLactationLipid metabolismEpithelial specificmRNAProteinProfiling

Abbreviations

ADPH

Adipophilin

ACLY

ATP Citrate Lyase

Csn2

β-casein

ChREBP

Carbohydrate Responsive Element Binding Protein

C/EBPβ and C/EBPδ

CCAAT/enhancer-binding protein beta and delta

Ker18, Ker19

Cytokeratin 18 and 19

LCM

Laser Capture Micro-dissection

MECs

Mammary Epithelial Cells

PLIN

Perilipin

qPCR

quantitative Real Time PCR

THRSP, Spot14

Thyroid Hormone Responsive Protein Spot 14

TAGs

triglycerides

WTLs

Whole mammary Tissue Lysates

Copyright information

© Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Michael C. Rudolph
    • 1
    • 2
  • Elizabeth A. Wellberg
    • 1
  • Steven M. Anderson
    • 1
    • 2
    • 3
  1. 1.Department of PathologyUniversity of Colorado DenverAuroraUSA
  2. 2.Program in Molecular BiologyUniversity of Colorado DenverAuroraUSA
  3. 3.School of Medicine, Department of PathologyUniversity of Colorado DenverAuroraUSA