Leukemic Stem Cell Culture in Cytokine-Free Medium

Methods Mol Biol. 2021:2185:259-265. doi: 10.1007/978-1-0716-0810-4_15.

Abstract

Leukemia-initiating cells, also known as leukemic stem cells (LSCs), are experimentally defined by their ability to engraft immunocompromised mice and are believed to be a major cause of relapse in acute myeloid leukemia (AML). Despite the aggressive characteristics of acute leukemia, AML blasts are difficult to culture once removed from the patient, and LSCs, which are a minor fraction of the blast population, are especially difficult to transplant after culture. This impedes development of new therapies for AML that target LSCs. Here, we present a simple strategy to culture LSCs in cytokine-free medium and to perform flow cytometric analysis of the resulting cell population for the characterization of LSCs maintenance and differentiation.

Keywords: Acute myeloid leukemia (AML); Cytokine-free culture medium; Flow cytometry; GSK-3; Leukemia-initiating cells; Leukemic stem cells; mTOR.

MeSH terms

  • Cell Culture Techniques*
  • Cytokines / pharmacology
  • Humans
  • Leukemia, Myeloid, Acute* / metabolism
  • Leukemia, Myeloid, Acute* / pathology
  • Neoplastic Stem Cells / metabolism
  • Neoplastic Stem Cells / pathology
  • Tumor Cells, Cultured

Substances

  • Cytokines