PhD Scientific Days 2025

Poster Session III. - C: Molecular Medicine

Improvement of Transient Agarose Spot (TAS) Cell Migration Assay: Microplate-based Detection and Evaluation

Name of the presenter

Veres-Székely Apor

Institute/workplace of the presenter

Pediatric Center, Semmelweis University, Budapest, Hungary; HUN-REN – SU Pediatrics and Nephrology Research Group, Budapest, Hungary

Authors

Apor Veres-Székely¹, Csenge Szász², Domonkos Pap¹, Péter Bokrossy², Dorina Lenzinger³, Tamás Visnovitz⁴, Judith Mihály⁵, Marcell Pálmai⁵, Zoltán Varga⁶, László Őrfi⁷, Attila J. Szabó¹, Ádám Vannay¹, Beáta Szebeni¹

1: Pediatric Center, Semmelweis University, Budapest, Hungary; HUN-REN – SU Pediatrics and Nephrology Research Group, Budapest, Hungary
2: Pediatric Center, Semmelweis University, Budapest, Hungary
3: Department of Genetics, Cell- and Immunobiology, Semmelweis University, Budapest, Hungary
4: Department of Genetics, Cell- and Immunobiology, Semmelweis University, Budapest, Hungary; Department of Plant Physiology and Molecular Plant Biology, Eötvös Loránd University, Budapest, Hungary
5: TTK Biological Nanochemistry Research Group, Institute of Materials and Environmental Chemistry, Research Centre for Natural Sciences, Budapest, Hungary
6: TTK Biological Nanochemistry Research Group, Institute of Materials and Environmental Chemistry, Research Centre for Natural Sciences, Budapest, Hungary; Department of Physical Chemistry and Materials Science, Faculty of Chemical Technology and Biotech-nology, Budapest University of Technology and Economics, Budapest, Hungary
7: Vichem Chemie Research Ltd., Budapest, Hungary

Text of the abstract

Introduction
Collective cell migration plays a vital role in various biological processes, such as tumor progression and metastasis. The conventional scratch assay (wound healing assay) is widely used to study this phenomenon, but it has significant limitations in terms of throughput, reproducibility, and data analysis.
Aims
To address the shortcomings of traditional migration assays, we aimed to further improve our previously developed Transient Agarose Spot (TAS) assay by adapting it to a high-throughput, microplate-based format suitable for automated analysis.
Methods
We established a 96-well plate version of the TAS assay and integrated it with microplate reader-based automated data acquisition. Hoechst staining was used to label viable cells, allowing for reliable, non-toxic kinetic measurements. Fluorophore-expressing cancer cells were employed to validate the method.
Results
The microplate-based TAS assay enabled accurate, high-throughput detection of cell migration. It effectively measured the dose-dependent influence of fetal bovine serum (FBS) on migration, as well as the anti-migratory effects of kinase inhibitors and mesenchymal stem cell-derived extracellular vesicles (EVs) in lung cancer cells. The assay provided quantifiable, concentration-dependent results, demonstrating both inhibitory and modulatory effects on cell migration.
Conclusion
This improved TAS assay offers a robust, scalable, and cost-effective alternative to traditional microscopy-based methods. Its compatibility with high-throughput screening makes it a valuable tool for studying cell migration and for drug discovery applications.
Funding
This research was supported by the HUN-REN-SE Pediatric and Nephrology Research Group, National Research, Development and Innovation Office (NKFIH) K-142728 (A.J.S.), TKP2021-EGA-24 (A.J.S.), TKP2021-EGA-23 (T.V.), EKÖP-2024-53 (A.V.); EKÖP-2024-160 (P.B.); EKÖP-2024-162 (C.S.); VEKOP-2.3.3-15-2017-00016 (T.V.), RRF-2.3.121-2022-00003 (T.V.), Hungarian Academy of Sciences, János Bolyai Research Scholarship (D.P., T.V., M.P.).