Migration Assay for Leukemic Cells in a 3D Matrix Toward a Chemoattractant

Methods Mol Biol. 2019;2017:97-107. doi: 10.1007/978-1-4939-9574-5_8.


In leukemia, leukemic cells hijack the hematopoietic stem cell (HSC) microenvironment in the bone marrow-the so-called stem cell niche-by flooding the niche with clonal progeny of leukemic cells. They can exploit signaling pathways which are critical for HSC development to support their own survival, homing, and maintenance. These interactions of leukemic cells with the microenvironment have an impact on therapy progress and patient outcome. Therefore, signals for homing and anchorage of leukemic cells to the bone marrow have to be investigated by using tools that allow the migration of cells toward critical signals. Here, we describe an in vitro migration assay for leukemic cells toward a chemoattractant in a 3D environment exemplified by migration of the cell line OCI-AML3 to a CXC motif chemokine ligand 12 (CXCL12) gradient. For this purpose, a chemotaxis slide is filled with a hydrogel system mimicking the extracellular matrix in vivo. The cells are encapsulated into the hydrogel network during polymerization, and a CXCL12 gradient is introduced in the enclosed chambers to trigger migration. Cell migration in the 3D network of the hydrogel is monitored by time-lapse microscopy. We describe the experimental setup and the tools for cell tracking and data analysis.

Keywords: 3D matrix; CXCR4/CXCL12 axis; Chemokine gradient; Leukemic cells; Migration; μ-Slides.

MeSH terms

  • Cell Line, Tumor / cytology*
  • Cell Line, Tumor / metabolism
  • Cell Movement
  • Cell Tracking
  • Chemokine CXCL12 / metabolism*
  • Chemotaxis
  • Humans
  • Hydrogels / chemistry
  • Leukemia / pathology*
  • Stem Cell Niche
  • Tumor Microenvironment


  • CXCL12 protein, human
  • Chemokine CXCL12
  • Hydrogels