Establishment and Characterization of Long-Term Cultures Derived from Primary Acute Myeloid Leukemia Cells for HDAC Inhibitor Research

Methods Mol Biol. 2017:1510:127-148. doi: 10.1007/978-1-4939-6527-4_10.

Abstract

Histone deacetylase (HDAC) inhibitors are promising drugs. These agents lead to growth inhibition, cell cycle arrest, premature senescence, and apoptosis of malignant cells. Aim of our studies was to determine the efficacy of HDAC inhibitors on the clinically most relevant population of human leukemic progenitor cells in vitro. We here present stroma-free long-term cultures (LTC) of primary acute myeloid leukemia (AML) cells as a useful system for drug sensitivity testing in functional assays. AML-LTC are established by isolating mononuclear cells from peripheral blood samples of AML patients followed by selection of CD34+ progenitor cells. AML-LTC cells can be maintained in liquid culture supplemented with cytokines and utilized for in vitro analyses to assess proliferation, apoptosis, expression of surface proteins or intracellular proteins and signal transduction, respectively.

Keywords: AML long-term culture; CD34+ progenitor cells; HDAC inhibitor; Leukemia.

MeSH terms

  • Antigens, CD34 / genetics
  • Antigens, CD34 / metabolism
  • Apoptosis / drug effects
  • Biomarkers, Tumor / genetics*
  • Biomarkers, Tumor / metabolism
  • Cell Culture Techniques*
  • Cell Proliferation / drug effects
  • Cell Separation / methods
  • Cytokines / pharmacology
  • Gene Expression Regulation, Neoplastic*
  • Histone Deacetylase Inhibitors / pharmacology*
  • Histone Deacetylases / genetics*
  • Histone Deacetylases / metabolism
  • Humans
  • Hydroxamic Acids / pharmacology
  • Indoles / pharmacology
  • Inhibitor of Apoptosis Proteins / genetics
  • Inhibitor of Apoptosis Proteins / metabolism
  • Leukemia, Myeloid, Acute / genetics
  • Leukemia, Myeloid, Acute / metabolism
  • Leukemia, Myeloid, Acute / pathology
  • Neoplastic Stem Cells / drug effects*
  • Neoplastic Stem Cells / metabolism
  • Neoplastic Stem Cells / pathology
  • Panobinostat
  • Primary Cell Culture
  • Signal Transduction
  • Survivin
  • Tumor Cells, Cultured
  • Valproic Acid / pharmacology
  • Vorinostat

Substances

  • Antigens, CD34
  • BIRC5 protein, human
  • Biomarkers, Tumor
  • Cytokines
  • Histone Deacetylase Inhibitors
  • Hydroxamic Acids
  • Indoles
  • Inhibitor of Apoptosis Proteins
  • Survivin
  • Vorinostat
  • Valproic Acid
  • Panobinostat
  • Histone Deacetylases