Cell cycle measurement of mouse hematopoietic stem/progenitor cells

Methods Mol Biol. 2014:1185:65-78. doi: 10.1007/978-1-4939-1133-2_5.

Abstract

Lifelong production of blood cells is sustained by hematopoietic stem cells (HSC). HSC reside in a mitotically quiescent state within specialized areas of the bone marrow (BM) microenvironment known as the hematopoietic niche (HN). HSC enter into active phases of cell cycle in response to intrinsic and extrinsic biological cues thereby undergoing differentiation or self-renewal divisions. Quiescent and mitotically active HSC have different metabolic states and different functional abilities such as engraftment and BM repopulating potential following their transplantation into conditioned recipients. Recent studies reveal that various cancers also utilize the same mechanisms of quiescence as normal stem cells and preserve the root of malignancy thus contributing to relapse and metastasis. Therefore, exploring the stem cell behavior and function in conjunction with their cell cycle status has significant clinical implications in HSC transplantation and in treating cancers. In this chapter, we describe methodologies to isolate or analytically measure the frequencies of quiescent (G0) and active (G1, S, and G2-M) hematopoietic progenitor and stem cells among murine BM cells.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Antigens, CD / metabolism
  • Benzimidazoles / metabolism
  • Biomarkers / metabolism
  • Bone Marrow Cells / cytology
  • Cell Cycle*
  • Cell Separation
  • Hematopoietic Stem Cells / cytology*
  • Hematopoietic Stem Cells / metabolism
  • Mice
  • Pyronine / metabolism
  • Receptors, Transferrin / metabolism
  • Staining and Labeling

Substances

  • Antigens, CD
  • Benzimidazoles
  • Biomarkers
  • CD71 antigen
  • Receptors, Transferrin
  • bisbenzimide ethoxide trihydrochloride
  • Pyronine