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@ARTICLE{Tomasova:863917,
author = {Tomasova, Lea and Guttenberg, Zeno and Hoffmann, Bernd and
Merkel, Rudolf},
title = {{A}dvanced 2{D}/3{D} cell migration assay for faster
evaluation of chemotaxis of slow-moving cells},
journal = {PLOS ONE},
volume = {14},
number = {7},
issn = {1932-6203},
address = {San Francisco, California, US},
publisher = {PLOS},
reportid = {FZJ-2019-03880},
pages = {e0219708 -},
year = {2019},
abstract = {Considering the essential role of chemotaxis of adherent,
slow-moving cells in processes such as tumor metastasis or
wound healing, a detailed understanding of the mechanisms
and cues that direct migration of cells through tissues is
highly desirable. The state-of-the-art chemotaxis
instruments (e.g. microfluidic-based devices, bridge assays)
can generate well-defined, long-term stable chemical
gradients, crucial for quantitative investigation of
chemotaxis in slow-moving cells. However, the majority of
chemotaxis tools are designed for the purpose of an
in-depth, but labor-intensive analysis of migratory behavior
of single cells. This is rather inefficient for applications
requiring higher experimental throughput, as it is the case
of e.g. clinical examinations, chemoattractant screening or
studies of the chemotaxis-related signaling pathways based
on subcellular perturbations. Here, we present an advanced
migration assay for accelerated and facilitated evaluation
of the chemotactic response of slow-moving cells. The
revised chemotaxis chamber contains a hydrogel
microstructure–the migration arena, designed to enable
identification of chemotactic behavior of a cell population
in respect to the end-point of the experiment. At the same
time, the assay in form of a microscopy slide enables direct
visualization of the cells in either 2D or 3D environment,
and provides a stable and linear gradient of
chemoattractant. We demonstrate the correctness of the assay
on the model study of HT-1080 chemotaxis in 3D and on 2D
surface. Finally, we apply the migration arena chemotaxis
assay to screen for a chemoattractant of primary
keratinocytes, cells that play a major role in wound
healing, being responsible for skin re-epithelialization and
a successful wound closure. In direction of new therapeutic
strategies to promote wound repair, we identified the
chemotactic activity of the epithelial growth factor
receptor (EGFR) ligands EGF and TGFα (transforming growth
factor α).},
cin = {ICS-7},
ddc = {610},
cid = {I:(DE-Juel1)ICS-7-20110106},
pnm = {552 - Engineering Cell Function (POF3-552)},
pid = {G:(DE-HGF)POF3-552},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:31314801},
UT = {WOS:000482331900039},
doi = {10.1371/journal.pone.0219708},
url = {https://juser.fz-juelich.de/record/863917},
}