Application of Concave Microwells to Pancreatic Tumor Spheroids Enabling Anticancer Drug Evaluation in a Clinically Relevant Drug Resistance Model
Open accessAbstract
Intrinsic drug resistance of pancreatic ductal adenocarcinoma (PDAC) warrants studies using models that are more clinically relevant for identifying novel resistance mechanisms as well as for drug development. Tumor spheroids (TS) mimic in vivo tumor conditions associated with multicellular resistance and represent a promising model for efficient drug screening, however, pancreatic cancer cells often fail to form spheroids using conventional methods such as liquid overlay. This study describes the induction of TS of human pancreatic cancer cells (Panc-1, Aspc-1, Capan-2) in concave polydimethylsiloxane (PDMS) microwell plates and evaluation of their usefulness as an anticancer efficacy test model. All three cell lines showed TS formation with varying degree of necrosis inside TS. Among these, Panc-1 spheroid with spherical morphology, a rather rough surface, and unique adhesion structures were successfully produced with no notable necrosis in concave microwell plates. Panc-1 TS contained growth factors or enzymes such as TGF-β1, CTGF, and MT1-MMP, and extracellular matrix proteins such as collagen type I, fibronectin, and laminin. Panc-1 cells grown as TS showed changes in stem cell populations and in expression levels of miRNAs that may play roles in chemoresistance. Visualization of drug penetration and detection of viability indicators, such as Ki-67 and MitoSOX, were optimized for TS for quantitative analysis. Water-soluble tetrazolium (MTS) and acid phosphatase (APH) assays were also successfully optimized. Overall, we demonstrated that concave PDMS microwell plates are a novel platform for preparation of TS of weakly aggregating cells and that Panc-1 spheroids may represent a novel three-dimensional model for anti-pancreatic cancer drug screening.
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Electron microscopy, including scanning and transmission modalities, provides ultra-high-resolution imaging and remains valuable for evaluating cellular morphology, surface topology, and intercellular organization across spheroid layers.
“Electron microscopy, including both scanning (SEM) and transmission (TEM) modalities, provides ultra-high-resolution imaging and remains a valuable tool for evaluating cellular morphology, surface topology, and intercellular organization across spheroid layers [245,256].”
From Modeling Tumor Microenvironment Complexity In Vitro: Spheroids as Physiologically Relevant Tumor Models and Strategies for Their Analysis (2025) · cites this paper as [245]
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- Authors
- Yeon, S.-E., No, D. Y., Lee, S.-H., Nam, S. W., Oh, I.-H., Lee, J., & Kuh, H.-J.
- Affiliations
- Catholic University of Korea (KR)
- Korea University (KR)
- Chung-Ang University (KR)
- Corresponding author
- Kuh, Hyo-Jeong
- Venue
- PLoS ONE
- Publisher
- Public Library of Science (PLoS)
- Published
- 2013-09-10
- Volume
- 8
- Issue
- 9
- Pages
- e73345
- ISSN
- 1932-6203 (ISSN-L)
- PMC
- PMC3769301
- OpenAlex
- W2025183054
- Cited by
- 110
- References
- 42
- Citations by year
- 2026: 1 · 2025: 6 · 2024: 6 · 2023: 4 · 2022: 4 · 2021: 12 · 2020: 14 · 2019: 12 · 2018: 11 · 2017: 13 · 2016: 11 · 2015: 9 · 2014: 6
- FWCI
- 4.79
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- 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)
- Funders
- National Research Foundation
- National Research Foundation of Korea (2012R1A5A2047939, 2012R1A2A2A01003361)
- Topics
- Physical Sciences › Engineering › Biomedical Engineering › 3D Printing in Biomedical Research
- Health Sciences › Medicine › Oncology › Cancer Cells and Metastasis
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular Mechanics and Interactions
- Keywords
- pancreatic ductal adenocarcinoma, drug resistance, anticancer drug screening, PANC-1 cells, 3D tumor models, extracellular matrix, chemoresistance, drug penetration, necrosis, stem cell populations, microRNA expression, MTS assay
- MeSH terms
- Antinematodal Agents, Drug Screening Assays, Antitumor, Equipment Design, Humans, Pancreatic Neoplasms, Spheroids, Cellular, Cell Line, Tumor
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