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Identification and Separation of Normal Hematopoietic Stem Cells and Leukemia Stem Cells from Patients with Acute Myeloid Leukemia

  • Van T. Hoang
  • Isabel Hoffmann
  • Karina Borowski
  • Abraham Zepeda-Moreno
  • Dan Ran
  • Eike C. Buss
  • Patrick Wuchter
  • Volker Eckstein
  • Anthony D. Ho
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1035)

Abstract

Mounting evidences indicate that leukemic cells in patients with acute myeloid leukemia (AML) are derived from leukemia stem cells (LSC). In analogy to normal hematopoietic stem cells (HSC), LSC remain mostly dormant and are hence resistant to conventional chemotherapy. Residual, physiological HSC exist alongside with LSC, with heterogeneous dominance of LSC over HSC in individual patients. We have devised a flow cytometric method for the identification and separation of these two stem cell populations based on surface antigen markers such as CD34, CD38, lineage aberrant markers, and aldehyde dehydrogenase (ALDH) enzyme activity.

Key words

Acute myeloid leukemia Leukemia stem cells Hematopoietic stem cells Flow cytometry FACS Aldehyde dehydrogenase Aberrant markers 

Abbreviations

ALDH

Aldehyde dehydrogenase

AM

Aberrant marker

AML

Acute myeloid leukemia

APC

Allophycocyanin

FSC

Forward scatter

FACS

Fluorescence-activated cell sorting

FITC

Fluorescein

HSC

Hematopoietic stem cell

LSC

Leukemia stem cells

MNC

Mononuclear cells

NOD/SCIDNOD/SCID

Non-Obese Diabetic/Severe Combined Immune Deficiency

NSG

NOD/SCID/interleukin 2 receptor gammanull

PE

Phycoerythrin

PI

Propidium iodide

SSC

Side scatter

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Copyright information

© Springer Science+Business Media, LLC 2013

Authors and Affiliations

  • Van T. Hoang
    • 1
  • Isabel Hoffmann
    • 1
  • Karina Borowski
    • 1
  • Abraham Zepeda-Moreno
    • 1
  • Dan Ran
    • 1
  • Eike C. Buss
    • 1
  • Patrick Wuchter
    • 1
  • Volker Eckstein
    • 1
  • Anthony D. Ho
    • 1
  1. 1.Department of Internal Medicine VUniversity of HeidelbergHeidelbergGermany

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