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Cited by 87Cell Image Analysis TechniquesCellular Mechanics and InteractionsCancer Cells and Metastasis

Quantification of dynamic morphological drug responses in 3D organotypic cell cultures by automated image analysis

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Härmä, Ville, Schukov, Hannu-Pekka, Happonen, Antti, Ahonen, Ilmari, Virtanen, Johannes, Siitari, Harri, Åkerfelt, Malin, Lötjönen, Jyrki +1 more·2014·PLoS ONE
DOI 10.1371/journal.pone.0096426PMC PMC4014501
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Abstract

Glandular epithelial cells differentiate into complex multicellular or acinar structures, when embedded in three-dimensional (3D) extracellular matrix. The spectrum of different multicellular morphologies formed in 3D is a sensitive indicator for the differentiation potential of normal, non-transformed cells compared to different stages of malignant progression. In addition, single cells or cell aggregates may actively invade the matrix, utilizing epithelial, mesenchymal or mixed modes of motility. Dynamic phenotypic changes involved in 3D tumor cell invasion are sensitive to specific small-molecule inhibitors that target the actin cytoskeleton. We have used a panel of inhibitors to demonstrate the power of automated image analysis as a phenotypic or morphometric readout in cell-based assays. We introduce a streamlined stand-alone software solution that supports large-scale high-content screens, based on complex and organotypic cultures. AMIDA (Automated Morphometric Image Data Analysis) allows quantitative measurements of large numbers of images and structures, with a multitude of different spheroid shapes, sizes, and textures. AMIDA supports an automated workflow, and can be combined with quality control and statistical tools for data interpretation and visualization. We have used a representative panel of 12 prostate and breast cancer lines that display a broad spectrum of different spheroid morphologies and modes of invasion, challenged by a library of 19 direct or indirect modulators of the actin cytoskeleton which induce systematic changes in spheroid morphology and differentiation versus invasion. These results were independently validated by 2D proliferation, apoptosis and cell motility assays. We identified three drugs that primarily attenuated the invasion and formation of invasive processes in 3D, without affecting proliferation or apoptosis. Two of these compounds block Rac signalling, one affects cellular cAMP/cGMP accumulation. Our approach supports the growing needs for user-friendly, straightforward solutions that facilitate large-scale, cell-based 3D assays in basic research, drug discovery, and target validation.

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  • An increased proportion of spheroids and invasive spheroids was observed in all tumor cell lines (PC3, DU145, LNCaP, WPE1-NB26, MCF7) compared with control cell lines (RWPE-1 and MCF10A).

    “As expected, we observed an increased proportion of spheroids and invasive spheroids in all tumor cell lines (PC3, DU145, LNCaP, WPE1-NB26 and MCF7) compared with the control cell lines (RWPE-1 and MCF10A) (30).”

    From Direct transfection of clonal organoids in Matrigel microbeads: a promising approach toward organoid-based genetic screens (2018) · cites this paper as [30]

    The Results (Experimental validation; page not provided) contradict the claim that all listed tumor lines had more invasive spheroids than the controls: LNCaP and DU145 typically formed round spheroids lacking invasive properties, while the control line RWPE-1 formed branching structures. The paper does not report the claimed comparison of spheroid proportions across these lines. (LLM, conf=0.88)

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  • In contrast with a previous study demonstrating low plasmid transfection efficiency (about 6%) in 3D cultures, the present system reached plasmid extinction efficiency of more than 40%.

    “In contrast with a previous study demonstrating low plasmid transfection efficiency (about 6%) in 3D cultures (30), we managed to reach plasmid extinction efficiency of more than 40%.”

    From Direct transfection of clonal organoids in Matrigel microbeads: a promising approach toward organoid-based genetic screens (2018) · cites this paper as [30]

    The citation is attached to the claim that a previous study found about 6% plasmid transfection efficiency in 3D cultures; the citing authors' own result of more than 40% is not attributed to this source. The Materials and Methods section describes 3D culture preparation, but neither it nor the retrieved passages report plasmid transfection efficiency (searched: 6% plasmid transfection, plasmid gene delivery). (LLM, conf=0.72)

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Research works printed but not linked: 53

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Authors
Härmä, V., Schukov, H.-P., Happonen, A., Ahonen, I., Virtanen, J., Siitari, H., Åkerfelt, M., Lötjönen, J., & Nees, M.
Affiliations
  • University of Turku (FI)
  • Tampere University of Technology (FI)
  • VTT Technical Research Centre of Finland (FI)
Corresponding author
Nees, Matthias
ORCID
Venue
PLoS ONE
Publisher
Public Library of Science (PLoS)
Published
2014-05-08
Volume
9
Issue
5
Pages
e96426
ISSN
1932-6203 (ISSN-L)
DOI
10.1371/journal.pone.0096426
PMC
PMC4014501
OpenAlex
W1985090392
Cited by
87
References
61
Citations by year
2026: 7 · 2025: 1 · 2024: 8 · 2023: 5 · 2022: 10 · 2021: 7 · 2020: 6 · 2019: 9 · 2018: 7 · 2017: 8 · 2016: 9 · 2015: 10
FWCI
5.36
Type
journal article
Language
en
Open access
Licence
Locations
  • PLoS ONE (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, other-oa)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • Figshare (submittedVersion, cc-by)
  • Åbo Akademi University Research Portal (submittedVersion, other-oa)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Cell Image Analysis Techniques
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular Mechanics and Interactions
  • Health Sciences › Medicine › Oncology › Cancer Cells and Metastasis
Keywords
tumor spheroids, tumor invasion, automated image analysis, morphometrics, high-content screening, prostate cancer, breast cancer, cell motility, drug discovery
MeSH terms
Cell Differentiation, Cell Movement, Epithelial Cells, Extracellular Matrix, Humans, Image Processing, Computer-Assisted, Cell Line, Tumor, Cell Shape, Cell Proliferation

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