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

  • Sabrina Zippel
  • Annamarija Raic
  • Cornelia Lee-ThedieckEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2017)


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.

Key words

Leukemic cells Migration 3D matrix μ-Slides CXCR4/CXCL12 axis Chemokine gradient 



Sabrina Zippel and Annamarija Raic contributed equally to this work.


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Sabrina Zippel
    • 1
    • 2
  • Annamarija Raic
    • 1
    • 2
  • Cornelia Lee-Thedieck
    • 1
    • 2
    Email author
  1. 1.Institute of Functional InterfacesKarlsruhe Institute of Technology (KIT)Eggenstein-LeopoldshafenGermany
  2. 2.Institute of Cell Biology and BiophysicsLeibniz University HannoverHannoverGermany

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