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Related Experiment Video

Updated: Mar 24, 2026

Quantification of Self-renewal in Murine Mammosphere Cultures
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Murine mammary stem/progenitor cell isolation: Different method matters?

Hui Gao1, Qiaoxiang Dong2, Yuanhong Chen3

  • 1School of Laboratory Medicine and Life Science, Wenzhou Medical University, University Town, Wenzhou, 325035 China ; Department of Cellular and Structural Biology, University of Texas Health Science Center, San Antonio, TX 78299 USA.

Springerplus
|March 3, 2016
PubMed
Summary

Comparing mammary stem cell isolation methods in mice, this study found slow overnight digestion superior to fast methods. CD49f/EpCAM markers best isolated basal cells, crucial for accurate gene expression analysis.

Keywords:
Digestion methodEnrichmentFlow cytometric analysisMammary stem/progenitor cellSurface marker

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Area of Science:

  • Cell Biology
  • Developmental Biology
  • Stem Cell Research

Background:

  • Murine mammary stem/progenitor cell isolation is vital for research.
  • Existing isolation methods lack direct comparative analysis.
  • Understanding optimal methods is key for reproducible stem cell research.

Purpose of the Study:

  • To compare the efficiency of two digestion methods and three surface marker sets for isolating murine mammary stem and progenitor cells.
  • To evaluate method performance in C57BL/6J and FVB mouse strains.
  • To identify the most reliable method for enriching mammary stem and progenitor cells.

Main Methods:

  • Compared two distinct enzymatic digestion protocols (slow overnight vs. fast methods).
  • Evaluated three surface marker combinations (CD24/CD29, CD49f/EpCAM, etc.) for cell sorting.
  • Assessed cell isolation efficiency and purity in C57BL/6J and FVB mice.

Main Results:

  • Slow overnight digestion using collagenase/hyaluronidase provided consistent and reliable enrichment of mammary stem/progenitor cells.
  • Fast digestion protocols resulted in high non-epithelial cell contamination and low basal cell yield.
  • CD49f/EpCAM marker set demonstrated the lowest non-epithelial cell inclusion in the basal cell gate, particularly in C57BL/6J mice.

Conclusions:

  • Gentle, slow overnight digestion is recommended for reproducible murine mammary stem/progenitor cell isolation.
  • The choice of surface markers significantly impacts basal cell purity, with CD49f/EpCAM being optimal.
  • Optimized isolation protocols are critical for accurate downstream analyses, such as gene expression profiling.