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Isolation and characterization of side population cells

Margaret A Goodell1, Shannon McKinney-Freeman, Fernando D Camargo

  • 1Center for Cell and Gene Therapy, Department of Immunology and Cell and Molecular Biology Program, Baylor College of Medicine, Houston, TX, USA.

Methods in Molecular Biology (Clifton, N.J.)
|September 14, 2004
PubMed

The protocol for isolation of side population (SP) cells was originally established for murine bone marrow hematopoietic stem cells (HSCs), but it has also been adapted for other species and tissues. This purification strategy offers a simple and reproducible strategy to obtain a highly homogeneous population of HSCs. The method is based on the differential efflux of the fluorescent DNA-binding dye Hoechst 33342 from stem cells relative to nonstem cells. The protocols outlined in this chapter describe the isolation of murine SP cells from both bone marrow and skeletal muscle using the fluorescent DNA-binding dye Hoechst 33342. In these tissues, the SP cells that are isolated are HSCs.

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