Hydrogel Microwell Arrays Allow the Assessment of Protease-Associated Enhancement of Cancer Cell Aggregation and Survival.
Current routine cell culture techniques are only poorly suited to capture the physiological complexity of tumor microenvironments, wherein tumor cell function is affected by intricate three-dimensional (3D), integrin-dependent cell-cell and cell-extracellular matrix (ECM) interactions. 3D cell cultures allow the investigation of cancer-associated proteases like kallikreins as they degrade ECM proteins and alter integrin signaling, promoting malignant cell behaviors. Here, we employed a hydrogel microwell array platform to probe using a high-throughput mode how ovarian cancer cell aggregates of defined size form and survive in response to the expression of kallikreins and treatment with paclitaxel, by performing microscopic, quantitative image, gene and protein analyses dependent on the varying microwell and aggregate sizes. Paclitaxel treatment increased aggregate formation and survival of kallikrein-expressing cancer cells and levels of integrins and integrin-related factors. Cancer cell aggregate formation was improved with increasing aggregate size, thereby reducing cell death and enhancing integrin expression upon paclitaxel treatment. Therefore, hydrogel microwell arrays are a powerful tool to screen the viability of cancer cell aggregates upon modulation of protease expression, integrin engagement and anti-cancer treatment providing a micro-scaled yet high-throughput technique to assess malignant progression and drug-resistance.
Faculty of Health, Institute of Health and Biomedical Innovation (IHBI), Queensland University of Technology (QUT), 60 Musk Avenue, Kelvin Grove 4059, Brisbane, Australia. daniela.lossner@qut.edu.au.
Daniela
D
Loessner
Laboratory of Stem Cell Bioengineering, Institute of Bioengineering, École Polytechnique Fédérale de Lausanne (EPFL), Building AI 3138, Station 15, CH-1015 Lausanne, Switzerland. stefan.kobel@roche.com.
Stefan
S
Kobel
Faculty of Health, Institute of Health and Biomedical Innovation (IHBI), Queensland University of Technology (QUT), 60 Musk Avenue, Kelvin Grove 4059, Brisbane, Australia. j.clements@qut.edu.au.
Judith A
JA
Clements
Laboratory of Stem Cell Bioengineering, Institute of Bioengineering, École Polytechnique Fédérale de Lausanne (EPFL), Building AI 3138, Station 15, CH-1015 Lausanne, Switzerland. matthias.lutolf@epfl.ch.
Matthias P
MP
Lutolf
Faculty of Science and Engineering, IHBI, QUT, 60 Musk Avenue, Kelvin Grove 4059, Brisbane, Australia. dietmar.hutmacher@qut.edu.au.
Dietmar W
DW
Hutmacher
bioengineered microenvironments; cell aggregates; integrins; kallikreins; microwell arrays; ovarian cancer; paclitaxel