Culture and Harvest of CpG-Stimulated Peripheral Blood or Bone Marrow in Chronic Lymphocytic Leukemia

Methods Mol Biol. 2019:1881:27-34. doi: 10.1007/978-1-4939-8876-1_3.

Abstract

Chromosome analysis of chronic lymphocytic leukemia (CLL) is an important clinical tool for evaluating prognosis and disease progression. Visualizing chromosomes microscopically using traditional cytogenetic techniques requires dividing cells to be arrested during metaphase. The major challenge for performing this analysis on CLL samples is stimulating the cells to divide in culture. Stimulation of CLL cells with CpG oligodeoxynucleotides has improved our ability to perform chromosome analysis for this leukemia. This protocol should help the reader successfully culture CLL samples for clinical chromosome analysis.

Keywords: B-cell mitogen; Chronic lymphocytic leukemia; CpG oligodeoxynucleotides; Cytogenetics; Karyotype.

MeSH terms

  • B-Lymphocytes / drug effects
  • Bone Marrow Cells / drug effects
  • Humans
  • Karyotyping / instrumentation
  • Karyotyping / methods
  • Leukemia, Lymphocytic, Chronic, B-Cell / blood
  • Leukemia, Lymphocytic, Chronic, B-Cell / genetics*
  • Leukemia, Lymphocytic, Chronic, B-Cell / pathology
  • Metaphase / drug effects
  • Mitogens / pharmacology
  • Oligodeoxyribonucleotides / pharmacology*
  • Primary Cell Culture / instrumentation
  • Primary Cell Culture / methods*
  • Specimen Handling / instrumentation
  • Specimen Handling / methods
  • Tumor Cells, Cultured

Substances

  • CPG-oligonucleotide
  • GNKG168
  • Mitogens
  • Oligodeoxyribonucleotides