Isolation and functional characterization of side population stem cells.
The "side population" (SP) phenotype is a manifestation of primitive cells' ability to efficiently efflux the fluorescent DNA-staining dye Hoechst 33342 and can be used as the basis by which to isolate these cells using flow cytometry. In the bone marrow (BM), the SP defines a cell subset with a highly homogeneous content of hematopoietic stem cells (HSCs). In this chapter, we describe a protocol to reproducibly isolate murine BM SP cells, as well as analytic measures, such as single cell transplantation, that can be used to assess the functionality of SP-derived stem cells.
Johnnidis
Jonathan B
JB
Whitehead Institute for Biomedical Research, Cambridge, MA, USA.
Camargo
Fernando D
FD
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology*; D002469:Cell Separation; D005434:Flow Cytometry; D051379:Mice; D008810:Mice, Inbred C57BL; D013234:Stem Cells / Q000166:cytology*