Protocol

Basic Cell Culture Protocols

Volume 946 of the series Methods in Molecular Biology pp 151-162

Date:

Isolation and Characterization of Mouse Side Population Cells

  • Aysegul V. ErgenAffiliated withThe Center for Cell and Gene Therapy, Baylor College of Medicine
  • , Mira JeongAffiliated withThe Center for Cell and Gene Therapy, Baylor College of MedicineUniversity of Science and Technology
  • , Kuanyin K. LinAffiliated withThe Center for Cell and Gene Therapy, Baylor College of Medicine
  • , Grant A. ChallenAffiliated withThe Center for Cell and Gene Therapy, Baylor College of Medicine
  • , Margaret A. GoodellAffiliated withStem Cells and Regenerative Medicine Center, Baylor College of Medicine Email author 

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Abstract

The side population (SP) is a subpopulation of mouse bone marrow cells highly enriched for hematopoietic stem cell activity. The SP is identified using flow cytometry as a minor population that efficiently effluxes the DNA-binding dye Hoechst 33342 relative to the rest of the bone marrow. Phenotypic and functionally analysis has established SP cells as highly phenotypically homogeneous and functional active. In this chapter we describe a detailed protocol for the purification of murine bone marrow SP cells based on Hoechst dye efflux in combination with the presence of HSC surface markers.

Key words

SP Hoechst 33342 Dye efflux Hematopoietic stem cell Purification Side population