title: Isolation and Assessment of Single Long-Term Reconstituting Hematopoietic Stem Cells from Adult Mouse Bone Marrow.
abstract: Hematopoietic stem cells with long-term repopulating activity can now be routinely obtained at purities of 40% to 50% from suspensions of adult mouse bone marrow. Here we describe robust protocols for both their isolation as CD45(+) EPCR(+) CD150(+) CD48(-) (ESLAM) cells using multiparameter cell sorting and for tracking their clonal growth and differentiation activity in irradiated mice transplanted with single ESLAM cells. The simplicity of these procedures makes them attractive for characterizing the molecular and biological properties of individual hematopoietic stem cells with unprecedented power and precision. © 2016 by John Wiley & Sons, Inc.
authors.affiliation: Wellcome Trust/MRC Stem Cell Institute, University of Cambridge, Cambridge, United Kingdom.|Department of Haematology, University of Cambridge, United Kingdom.
authors.forename: David G
authors.initials: DG
authors.lastname: Kent
authors.affiliation: Department of Dermatology, Brigham and Women's Hospital, Boston, Massachusetts.|Harvard Medical School, Boston, Massachusetts.
authors.forename: Brad J
authors.initials: BJ
authors.lastname: Dykstra
authors.affiliation: Terry Fox Laboratory, BC Cancer Agency, Vancouver, Canada.|Department of Medical Genetics, University of British Columbia, Vancouver, Canada.
authors.forename: Connie J
authors.initials: CJ
authors.lastname: Eaves
mesh_terms: D053687:Adult Stem Cells / Q000166:cytology*; D000375:Aging / Q000502:physiology*; D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology*; D002469:Cell Separation / Q000379:methods*; D058948:Cell Tracking; D005434:Flow Cytometry; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D007962:Leukocytes / Q000166:cytology; D008810:Mice, Inbred C57BL; D012038:Regeneration; D013997:Time Factors
keywords: flow cytometry; hematopoiesis; multi-lineage reconstitution; single-cell transplants; stem cell isolation