Ultrastructure and Function in In Vitro Cancer Models: From Morphology to Mechanism

A special issue of Biology (ISSN 2079-7737). This special issue belongs to the section "Cancer Biology".

Deadline for manuscript submissions: 15 July 2026 | Viewed by 592

Special Issue Editors


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Guest Editor
Department of Anatomy, Histology, Forensic Medicine and Orthopaedics, Section of Human Anatomy, Electron Microscopy Unit, University of Rome Sapienza, Via Alfonso Borelli 50, 00161 Rome, Italy
Interests: scanning electron microscopy; transmission electron microscopy
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Guest Editor
DIMES, Department of Experimental Medicine, Human Anatomy, Cellular Electron Microscopy Lab, University of Genoa, Genoa, Italy
Interests: breast cancers; targeted therapies; cell biology; endocytosis and trafficking; autophagy; electron microscopy; light microscopy; HER2/ERBB2 receptor; lysosomes
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

This Special Issue aims to bridge a critical gap by applying ultrastructural insights to functionally validate and mechanistically decode in vitro cancer models. While the existing literature often emphasizes genetic or molecular profiling, this Special Issue will highlight how high-resolution imaging moves beyond descriptive morphology to provide mechanistic explanations for phenomena like tumor heterogeneity, stromal crosstalk, and therapy response. We welcome studies that correlate structural alterations with functional phenotypes, thereby offering a complementary perspective that is essential for comprehensive and validated cancer modeling.

We are pleased to invite you to contribute to the Special Issue titled "Ultrastructure and Function in In Vitro Cancer Models: From Morphology to Mechanism", hosted in Biology (MDPI). This Special Issue aims to explore the ultrastructural features of cancer in vitro models, including organoids, spheroids, and other advanced 3D culture systems, using high-resolution imaging techniques such as Transmission Electron Microscopy (TEM), Scanning Electron Microscopy (SEM), Focused Ion Beam (FIB), and related modalities.

The focus of this Special Issue lies in enhancing our understanding of the subcellular and nanostructural hallmarks of cancer, as revealed by in vitro models. The scope includes both technical advancements and biological insights, encouraging submissions that combine morphological characterization with functional interpretation, particularly within tumor heterogeneity, cancer–stroma interaction, and therapy response studies.

By collecting original research, reviews, and methodological papers, this Special Issue seeks to supplement the current literature by showcasing how ultrastructural analysis can contribute to model validation, disease modeling, and therapeutic assessment. We welcome contributions that push the boundaries of electron microscopy and related techniques to shed light on the architectural complexity of cancer in vitro.

Dr. Michela Relucenti
Dr. Katia Cortese
Guest Editors

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Keywords

  • cancer
  • organoids
  • spheroids
  • transmission electron microscopy
  • scanning electron microscopy
  • focused ion beam (FIB)

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Published Papers (1 paper)

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Research

17 pages, 5897 KB  
Article
3D Breast Cancer Spheroids Reveal Architecture-Dependent HER2 Expression and Signaling
by Pietro Arnaldi, Valentina Delli Zotti, Grazia Bellese, Maria Cristina Gagliani, Paola Orecchia, Patrizio Castagnola and Katia Cortese
Biology 2025, 14(12), 1654; https://doi.org/10.3390/biology14121654 - 24 Nov 2025
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Abstract
Background: Three-dimensional (3D) culture systems offer a physiologically relevant alternative to monolayers for studying tumor organization, signaling, and drug response. HER2-positive breast cancers (BCa) account for 15–30% of BCa cases and benefit from HER2-targeted therapies, yet predictive in vitro models remain limited. Objective: [...] Read more.
Background: Three-dimensional (3D) culture systems offer a physiologically relevant alternative to monolayers for studying tumor organization, signaling, and drug response. HER2-positive breast cancers (BCa) account for 15–30% of BCa cases and benefit from HER2-targeted therapies, yet predictive in vitro models remain limited. Objective: To generate and compare 3D spheroids from two HER2+ BCa cell lines, SKBR3 and BT474, and investigate how 3D architecture influences HER2 distribution, intracellular signaling, and cellular organization. Methods: Spheroids were reproducibly generated from SKBR3 and BT474 cells and analyzed after 4 days of culture. Cell viability was evaluated using live/dead staining, HER2 distribution was assessed by confocal microscopy and quantified on cryosections, and protein expression/phosphorylation was measured by Western blotting. Epithelial and EMT markers were visualized by immunofluorescence, and ultrastructural features were examined by transmission electron microscopy (TEM). Results: Both cell lines formed viable spheroids with distinct architectures: SKBR3 spheroids were loose and heterogeneous, whereas BT474 spheroids were compact and highly spherical. Confocal and cryosection imaging showed consistent membrane HER2 localization with a progressive signal decrease toward the core of the spheroids, more pronounced in BT474. Western blotting revealed divergent HER2 expression and AKT phosphorylation: SKBR3 spheroids displayed increased HER2 but reduced pAKT, while BT474 spheroids showed reduced HER2 and pAKT levels. EpCAM and E-cadherin staining revealed cell line-specific epithelial organization, and TEM demonstrated differences in intercellular spacing and mitochondrial morphology, reflecting spheroid compactness. Conclusions: 3D architecture profoundly influences HER2 distribution, signaling, and structural organization in HER2+ BCa spheroids. This model provides a robust platform for investigating architecture-dependent molecular processes, with potential applications in drug response, receptor trafficking, and targeted therapy evaluation. Full article
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