Murine mammary stem/progenitor cell isolation: Different method matters?

Hui Gao, Qiaoxiang Dong, Yuanhong Chen, Fuchuang Zhang, Anqi Wu, Yuanshuo Shi, Abhik Bandyopadhyay, Benjamin J. Daniel, Changjiang Huang, Lu Zhe Sun

Research output: Contribution to journalArticle

3 Scopus citations

Abstract

Murine mammary stem/progenitor cell isolation has been routinely used in many laboratories, yet direct comparison among different methods is lacking. In this study, we compared two frequently used digestion methods and three sets of frequently used surface markers for their efficiency in enriching mammary stem and progenitor cells in two commonly used mouse strains, C57BL/6J and FVB. Our findings revealed that the slow overnight digestion method using gentle collagenase/hyaluronidase could be easily adopted and yielded reliable and consistent results in different batches of animals. In contrast, the different fast digestion protocols, as described in published studies, yielded high percent of non-epithelial cells with very few basal epithelial cells liberated in our hands. The three sets of markers tested in our hands reveal rather equally efficiency in separating luminal and basal cells if same fluorochrome conjugations were used. However, the tendency of non-epithelial cell inclusion in the basal cell gate was highest in samples profiled by CD24/CD29 and lowest in samples profiled by CD49f/EpCAM, this is especially true in mammary cells isolated from C57BL/6J mice. This finding will have significant implication when sorted basal cells are used for subsequent gene expression analysis.

Original languageEnglish (US)
Article number140
Pages (from-to)1-5
Number of pages5
JournalSpringerPlus
Volume5
Issue number1
DOIs
StatePublished - Dec 1 2016

Keywords

  • Digestion method
  • Enrichment
  • Flow cytometric analysis
  • Mammary stem/progenitor cell
  • Surface marker

ASJC Scopus subject areas

  • General

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