Migration Assay for Leukemic Cells in a 3D Matrix Toward a Chemoattractant
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 wordsLeukemic cells Migration 3D matrix μ-Slides CXCR4/CXCL12 axis Chemokine gradient
Sabrina Zippel and Annamarija Raic contributed equally to this work.
- 6.Kitaori T, Ito H, Schwarz EM, Tsutsumi R, Yoshitomi H, Oishi S, Nakano M, Fujii N, Nagasawa T, Nakamura T (2009) Stromal cell–derived factor 1/CXCR4 signaling is critical for the recruitment of mesenchymal stem cells to the fracture site during skeletal repair in a mouse model. Arthritis Rheumatol 60(3):813–823. https://doi.org/10.1002/art.24330CrossRefGoogle Scholar
- 7.Aiuti A, Webb IJ, Bleul C, Springer T, Gutierrez-Ramos JC (1997) The chemokine SDF-1 is a chemoattractant for human CD34+ hematopoietic progenitor cells and provides a new mechanism to explain the mobilization of CD34+ progenitors to peripheral blood. J Exp Med 185(1):111–120. https://doi.org/10.1084/jem.185.1.111CrossRefPubMedPubMedCentralGoogle Scholar
- 8.Cummins TD, Wu KZL, Bozatzi P, Dingwell KS, Macartney TJ, Wood NT, Varghese J, Gourlay R, Campbell DG, Prescott A, Griffis E, Smith JC, Sapkota GP (2018) PAWS1 controls cytoskeletal dynamics and cell migration through association with the SH3 adaptor CD2AP. J Cell Sci 131(1):12. https://doi.org/10.1242/jcs.202390CrossRefGoogle Scholar