title: CD201 and CD27 identify hematopoietic stem and progenitor cells across multiple murine strains independently of Kit and Sca-1.
abstract: Identification and isolation of hematopoietic stem cells (HSCs) in mice is most commonly based on the expression of surface molecules Kit and Sca-1 and the absence of markers of mature lineages. However, Sca-1 is absent or weakly expressed in hematopoietic progenitors in many strains, including nonobese diabetic (NOD), BALB/c, C3H, and CBA mice. In addition, both Kit and Sca-1 levels are modulated following bone marrow injury. In these cases, other markers and dye exclusion methods have been employed to identify HSCs, yet there is no antibody-based stain that enables identification of HSCs and early progenitors when Kit and Sca-1 are inadequate. CD201 is a marker that is highly restricted to HSCs and progenitors, and CD27 is expressed at moderate-to-high levels on HSCs. We show here that combining CD201 and CD27 enables highly efficient isolation of long-term HSCs in NOD mice as well as in other strains, including SJL, FVB, AKR, BALB/c, C3H, and CBA. We also find that HSCs appear to maintain expression of CD201 and CD27 after hematopoietic injury when Kit expression is downregulated. These results suggest a widely applicable yet simple alternative for HSC isolation in settings where Kit and Sca-1 expression are insufficient.
authors.affiliation: Department of Immunobiology, Joslin Diabetes Center, Boston, MA, USA; Harvard Medical School, Boston, MA, USA.
authors.forename: Sara E
authors.initials: SE
authors.lastname: Vazquez
authors.affiliation: Sue and Bill Gross Stem Cell Research Center, Department of Molecular Biology and Biochemistry, University of California Irvine, Irvine, CA, USA.
authors.forename: Matthew A
authors.initials: MA
authors.lastname: Inlay
authors.affiliation: Department of Immunobiology, Joslin Diabetes Center, Boston, MA, USA; Harvard Medical School, Boston, MA, USA. Electronic address: thomas.serwold@joslin.harvard.edu.
authors.forename: Thomas
authors.initials: T
authors.lastname: Serwold
mesh_terms: D000818:Animals; D000950:Antigens, Ly / Q000235:genetics / Q000502:physiology; D015551:Autoimmunity; D001773:Blood Cells / Q000737:chemistry* / Q000166:cytology; D001853:Bone Marrow / Q000528:radiation effects; D019070:Cell Lineage; D002469:Cell Separation / Q000379:methods*; D003922:Diabetes Mellitus, Type 1 / Q000097:blood / Q000235:genetics / Q000473:pathology; D000074283:Endothelial Protein C Receptor; D005434:Flow Cytometry / Q000379:methods*; D015870:Gene Expression; D006412:Hematopoietic Stem Cells / Q000737:chemistry* / Q000166:cytology; D008565:Membrane Proteins / Q000172:deficiency / Q000235:genetics / Q000502:physiology; D051379:Mice; D016688:Mice, Inbred NOD; D008815:Mice, Inbred Strains / Q000097:blood* / Q000235:genetics; D008822:Mice, Transgenic; D019009:Proto-Oncogene Proteins c-kit / Q000172:deficiency / Q000235:genetics / Q000502:physiology; D011828:Radiation Chimera; D011833:Radiation Injuries, Experimental / Q000097:blood / Q000473:pathology; D011956:Receptors, Cell Surface / Q000096:biosynthesis / Q000097:blood* / Q000235:genetics; D018127:Tumor Necrosis Factor Receptor Superfamily, Member 7 / Q000096:biosynthesis / Q000097:blood* / Q000235:genetics
chemical_list: D000950:Antigens, Ly; D000074283:Endothelial Protein C Receptor; C074650:Ly6a protein, mouse; D008565:Membrane Proteins; C080248:Procr protein, mouse; D011956:Receptors, Cell Surface; D018127:Tumor Necrosis Factor Receptor Superfamily, Member 7; D019009:Proto-Oncogene Proteins c-kit