Details
| Original language | English |
|---|---|
| Title of host publication | Stem Cell Mobilization |
| Subtitle of host publication | Methods and Protocols |
| Publisher | Humana Press |
| Pages | 97-107 |
| Number of pages | 11 |
| Edition | 1 |
| ISBN (electronic) | 978-1-4939-9574-5 |
| ISBN (print) | 978-1-4939-9573-8, 978-1-4939-9576-9 |
| Publication status | Published - 14 Jun 2019 |
Publication series
| Name | Methods in Molecular Biology |
|---|---|
| Volume | 2017 |
| ISSN (Print) | 1064-3745 |
| ISSN (electronic) | 1940-6029 |
Abstract
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
- Leukemic cells, Migration, 3D matrix, mu-Slides, CXCR4/CXCL12 axis, Chemokine gradient, μ-Slides
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Genetics
- Biochemistry, Genetics and Molecular Biology(all)
- Molecular Biology
Cite this
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Stem Cell Mobilization: Methods and Protocols. 1. ed. Humana Press, 2019. p. 97-107 (Methods in Molecular Biology; Vol. 2017).
Research output: Chapter in book/report/conference proceeding › Contribution to book/anthology › Research › peer review
}
TY - CHAP
T1 - Migration Assay for Leukemic Cells in a 3D Matrix Toward a Chemoattractant
AU - Zippel, Sabrina
AU - Raic, Annamarija
AU - Lee-Thedieck, Cornelia
PY - 2019/6/14
Y1 - 2019/6/14
N2 - 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.
AB - 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.
KW - Leukemic cells
KW - Migration
KW - 3D matrix
KW - mu-Slides
KW - CXCR4/CXCL12 axis
KW - Chemokine gradient
KW - μ-Slides
UR - http://www.scopus.com/inward/record.url?scp=85067521033&partnerID=8YFLogxK
U2 - 10.1007/978-1-4939-9574-5_8
DO - 10.1007/978-1-4939-9574-5_8
M3 - Contribution to book/anthology
SN - 978-1-4939-9573-8
SN - 978-1-4939-9576-9
T3 - Methods in Molecular Biology
SP - 97
EP - 107
BT - Stem Cell Mobilization
PB - Humana Press
ER -