CD201 and CD27 identify hematopoietic stem and progenitor cells across multiple murine strains independently of Kit and Sca-1

Exp Hematol. 2015 Jul;43(7):578-85. doi: 10.1016/j.exphem.2015.04.001. Epub 2015 Apr 16.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, Ly / genetics
  • Antigens, Ly / physiology
  • Autoimmunity
  • Blood Cells / chemistry*
  • Blood Cells / cytology
  • Bone Marrow / radiation effects
  • Cell Lineage
  • Cell Separation / methods*
  • Diabetes Mellitus, Type 1 / blood
  • Diabetes Mellitus, Type 1 / genetics
  • Diabetes Mellitus, Type 1 / pathology
  • Endothelial Protein C Receptor
  • Flow Cytometry / methods*
  • Gene Expression
  • Hematopoietic Stem Cells / chemistry*
  • Hematopoietic Stem Cells / cytology
  • Membrane Proteins / deficiency
  • Membrane Proteins / genetics
  • Membrane Proteins / physiology
  • Mice
  • Mice, Inbred NOD
  • Mice, Inbred Strains / blood*
  • Mice, Inbred Strains / genetics
  • Mice, Transgenic
  • Proto-Oncogene Proteins c-kit / deficiency
  • Proto-Oncogene Proteins c-kit / genetics
  • Proto-Oncogene Proteins c-kit / physiology
  • Radiation Chimera
  • Radiation Injuries, Experimental / blood
  • Radiation Injuries, Experimental / pathology
  • Receptors, Cell Surface / biosynthesis
  • Receptors, Cell Surface / blood*
  • Receptors, Cell Surface / genetics
  • Tumor Necrosis Factor Receptor Superfamily, Member 7 / biosynthesis
  • Tumor Necrosis Factor Receptor Superfamily, Member 7 / blood*
  • Tumor Necrosis Factor Receptor Superfamily, Member 7 / genetics

Substances

  • Antigens, Ly
  • Endothelial Protein C Receptor
  • Ly6a protein, mouse
  • Membrane Proteins
  • Procr protein, mouse
  • Receptors, Cell Surface
  • Tumor Necrosis Factor Receptor Superfamily, Member 7
  • Proto-Oncogene Proteins c-kit