Measuring ATP Concentration in a Small Number of Murine Hematopoietic Stem Cells

Methods Mol Biol. 2016:1516:205-217. doi: 10.1007/7651_2016_358.

Abstract

The metabolism of quiescent adult stem cells differs from the metabolism of differentiated cells. The metabolic processes are tightly regulated and their alterations disturb function of stem cells. One of the indicators of metabolic status of cells is the ATP level. While the method of measuring the ATP levels has been known for many years, estimating ATP levels in small population of defined stem cells isolated directly from the tissue has remained challenging. Here, we show our method of measuring the ATP levels in hematopoietic stem cells sorted from murine bone marrow. We used magnetic sorting as well as cell sorter and adopted the commonly used bioluminescence-based detection kits in described protocol. Our strategy allows to measure ATP levels in 1000 highly purified HSC.

Keywords: ATP; Cell sorting; Hematopoietic stem cells; Metabolism.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / isolation & purification*
  • Adenosine Triphosphate / metabolism
  • Animals
  • Bone Marrow Cells / metabolism*
  • Cell Differentiation / genetics
  • Flow Cytometry / methods*
  • Hematopoietic Stem Cells / cytology
  • Hematopoietic Stem Cells / metabolism*
  • Mice

Substances

  • Adenosine Triphosphate