Growing Role of 3D In Vitro Cell Cultures in the Study of Cellular and Molecular Mechanisms: Short Focus on Breast Cancer, Endometriosis, Liver and Infectious Diseases

Cells, Journal Year: 2024, Volume and Issue: 13(12), P. 1054 - 1054

Published: June 18, 2024

Over the past decade, development of three-dimensional (3D) models has increased exponentially, facilitating unravelling fundamental and essential cellular mechanisms by which cells communicate with each other, assemble into tissues organs respond to biochemical biophysical stimuli under both physiological pathological conditions. This section presents a concise overview most recent updates on significant contribution different types 3D cell cultures including spheroids, organoids organ-on-chip bio-printed in advancing our understanding molecular mechanisms. The case studies presented include breast cancer (BC), endometriosis, liver microenvironment infections. In BC, establishment culture permitted visualization role cancer-associated fibroblasts delivery exosomes, as well significance physical properties extracellular matrix promoting proliferation invasion. approach also become valuable tool gaining insight general specific drug resistance. Given considerable heterogeneity offer more accurate representation vivo microenvironment, thereby identification translation novel targeted therapeutic strategies. advantages provided hepatic environment, conjunction high throughput characterizing various platforms, have enabled elucidation complex underlying threatening diseases. A limited number for gut skin infections been developed. However, profound comprehension spatial temporal interactions between microbes, host their environment may facilitate advancement vitro, ex disease models. Additionally, it pave way approaches diverse research fields. interested reader will find concluding remarks challenges prospects using discovering areas covered this review.

Language: Английский

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Gravitational Forces and Matrix Stiffness Modulate the Invasiveness of Breast Cancer Cells in Bioprinted Spheroids

Materials Today Bio, Journal Year: 2025, Volume and Issue: 31, P. 101640 - 101640

Published: March 4, 2025

The progression of breast cancer is influenced by the stiffness extracellular matrix (ECM), which becomes stiffer as advances due to increased collagen IV and laminin secretion cancer-associated fibroblasts. Intriguingly, cells cultivated in two-dimensions exhibit a less aggressive behavior when exposed weightlessness, or microgravity conditions. This study aims elucidate interplay between on progression. For this purpose, three-dimensional spheroids cell lines (MCF-7 MDA-MB-231) were formed. These subsequently bioprinted hydrogels varying stiffness, obtained mixing gelatin methacrylate poly(ethylene) glycol diacrylate mixed at different ratios. constructs printed with custom stereolithography (SLA) bioprinter converted from low-cost, commercially available 3D printer. structures, encapsulating spheroids, then placed clinostat (microgravity simulation device) for duration seven days. Comparative analyses conducted objects cultured under standard earth gravity Protein expression was characterized through fluorescent microscopy, while gene MCF-7 analyzed via RNA sequencing. Remarkably, influence ECM protein levels could be modulated sometimes even reversed study's findings hold implications refining therapeutic strategies advanced stages - an array genes involved reversing metastatic might lead discovery new compounds that used clinical setting.

Language: Английский

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Extracellular Matrix Stiffness: Mechanotransduction and Mechanobiological Response-Driven Strategies for Biomedical Applications Targeting Fibroblast Inflammation

Polymers, Journal Year: 2025, Volume and Issue: 17(6), P. 822 - 822

Published: March 20, 2025

The extracellular matrix (ECM) is a dynamic network providing mechanical and biochemical cues that regulate cellular behavior. ECM stiffness critically influences fibroblasts, the primary producers, particularly in inflammation fibrosis. This review explores role of fibroblast-driven tissue remodeling, focusing on physicochemical biological mechanisms involved. Engineered materials, hydrogels, polydimethylsiloxane (PDMS) are highlighted for replicating tissue-specific stiffness, enabling precise control over cell–matrix interactions. surface functionalization substrate including collagen, polydopamine, fibronectin, enhances bioactivity fibroblast adhesion. Key mechanotransduction pathways, such as integrin signaling YAP/TAZ activation, related to regulating behaviors inflammatory responses. fibroblasts driving chronic diseases emphasizes their therapeutic potentials. Advances ECM-modifying strategies, tunable biomaterials hydrogel-based therapies, explored applications engineering, drug delivery, anti-inflammatory treatments, diagnostic tools accurate diagnosis prognosis stiffness-related diseases. integrates mechanobiology with biomedical innovations, comprehensive responses outlining future directions targeted therapies.

Language: Английский

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Quantitative characterization of the 3D self-organization of PDAC tumor spheroids reveals cell type and matrix dependence through advanced microscopy analysis

APL Bioengineering, Journal Year: 2025, Volume and Issue: 9(1)

Published: March 1, 2025

Pancreatic ductal adenocarcinoma (PDAC) is characterized by an abundant tumor-associated stroma composed from pancreatic stellate cells, which play a critical role in tumor progression. Developing accurate vitro models requires understanding the complex interactions between cells and their microenvironment. In this study, we present quantitative imaging-based characterization of three dimensional (3D) self-organization PDAC tumour spheroids using microfluidic platform that mimics key aspects Our model incorporates collagen type I hydrogels to recreate extracellular matrix, activated human (HPSCs), various cell types. Advanced imaging techniques, including Lattice Lightsheet Microscopy, allowed us analyze 3D growth spatial organization spheroids, revealing intricate biomechanical interactions. results indicate alterations matrix properties-such as stiffness, pore size, hydraulic permeability-due variations concentration significantly influence patterns depending on subtype epithelial-mesenchymal phenotype. Higher concentrations promoted larger epithelial-like lines, while mesenchymal-type required increased for into smaller spheroids. Furthermore, coculture with HPSCs affected spheroid formation distinctly based each line's genetic phenotypic traits. had opposing effects lines: one line exhibited enhanced growth, another showed inhibited formation, whereas mesenchymal-like minimal impact. These provide insights tumor-stroma interactions, emphasizing importance cell-specific matrix-dependent factors advancing our progression informing future therapeutic strategies.

Language: Английский

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A hybrid computational model of cancer spheroid growth with ribose-induced collagen stiffening

Frontiers in Bioengineering and Biotechnology, Journal Year: 2025, Volume and Issue: 13

Published: April 9, 2025

Metastasis, the leading cause of death in cancer patients, arises when cells disseminate from a primary solid tumour to distant organs. Growth and invasion often involve collective cell migration, which is profoundly influenced by cell-cell interactions extracellular matrix (ECM). The ECM's biochemical composition mechanical properties, such as stiffness, regulate behaviour migration dynamics. Mathematical modelling serves pivotal tool for studying predicting these complex dynamics, with hybrid discrete-continuous models offering powerful approach combining agent-based representations continuum descriptions surrounding microenvironment. In this study, we investigate impact ECM modulated via ribose-induced collagen cross-linking, on spheroid growth invasion. We employed model implemented PhysiCell simulate successfully replicating three-dimensional vitro experiments. incorporates detailed cell-ECM interactions, remodelling, proliferation. Our simulations align experimental observations two breast lines, non-invasive MCF7 invasive HCC 1954, under varying stiffness conditions. results demonstrate that increased due cross-linking inhibits cells, whereas remain largely unaffected. Furthermore, our show higher degradation not only enables but also facilitates formation multicellular protrusions. Conversely, increasing maximum speed can reach enhances while promoting single-cell This understanding interplay between proliferation, paving way future studies incorporating additional characteristics microenvironmental

Language: Английский

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An animal-free bioprocess to synthesize 3D human matrix scaffolds using mesenchymal stromal cells

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2025, Volume and Issue: unknown

Published: April 23, 2025

Abstract Basement membrane, the specialized extracellular matrix (ECM) that compartmentalizes endothelial and epithelial cells, is final frontier in bioengineering. The fibrillar collagens (Col I Col III) of interstitial are commonly used for tissue modeling, but networking scaffold basement membrane (human IV VI) remain elusive. Commercial hydrogels like Matrigel simulate ill-defined, dilute, variable contain murine proteins. Here, we investigated whether human mesenchymal stromal cells (MSCs) could be to produce 3D scaffolds amenable decellularization downstream applications Using a xeno-free, process, MSCs from placenta, umbilical cord, bone marrow adipose tissues were cultivated as adherent multilayers or free-floating spheroids. Matrix assembly was assessed daily by fluorescence imaging western blot, revealing de novo, systematic, development. Notably, these rich membrane-specific components VI. Interestingly, observed distinct combinations alpha chains other depending on their origin. Ongoing work aims combine this intrinsic patterning with strategic process parameters refine composition resemble target tissues, develop assays processes angiogenesis incorporate relevant biology readout.

Language: Английский

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Defining and modeling dynamic spatial heterogeneity within tumor microenvironments

Current Opinion in Cell Biology, Journal Year: 2024, Volume and Issue: 90, P. 102422 - 102422

Published: Aug. 30, 2024

Many solid tumors exhibit significant genetic, cellular, and biophysical heterogeneity which dynamically evolves during disease progression after treatment. This constant flux in cell composition, phenotype, spatial relationships, tissue properties poses challenges accurately diagnosing treating patients. Much of the complexity lies unraveling molecular changes different tumor compartments, how they influence one another space time where vulnerabilities exist that might be appropriate to target therapeutically. Recent advances profiling tools technologies are enabling new insight into underlying biology complex tumors, creating a greater understanding intricate relationship between types, states, microenvironment. Here we reflect on some recent discoveries this area, key knowledge technology gaps lie, advancements measurements vitro models for study intratumoral heterogeneity.

Language: Английский

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A Multiparametric Analysis Reveals Differential Behavior of Spheroid Cultures on Distinct Ultra-Low Attachment Plates Types

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: March 29, 2024

Abstract Spheroids have become principal three-dimensional biological models to study cancer, developmental processes, and drug efficacy. For spheroid generation, ultra-low attachment plates are noteworthy due their simplicity, compatibility with automation, experimental commercial accessibility. Nonetheless, it is unknown whether what degree the plate type impacts formation biology. This employed automated brightfield microscopy systematically compare size eccentricity of spheroids formed in six different types using four distinct human cell lines, i.e., CCD-1137Sk fibroblasts, HaCaT keratinocytes, MDA-MB-231 HT-29 cancer cells. Results showed that all exhibited similar sphe-roid-forming capabilities, gross patterns growth or shrinkage during days after seeding were comparable. Yet, varied among specific lines types. A confocal wholemount analysis by a novel pipeline AI-based 3D-image procedures revealed changes proliferation, number, nuclear volume, keratino-cyte differentiation, which accompanied altered YAP1-signals. The findings show may influence outcome campaigns. It advisable scan for optimal configuration investigation instead one standard kinds applications.

Language: Английский

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Multimodal Phasor Approach to study breast cancer cells invasion in 3D spheroid model

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: June 12, 2024

We implemented a multimodal set of functional imaging techniques optimized for deep-tissue to investigate how cancer cells invade surrounding tissues and their physiological properties change in the process. As model invasion extracellular matrix, we created 3D spheroids from triple-negative breast (MDA-MB-231) non-tumorigenic epithelial (MCF-10A). analyzed multiple hallmarks within same spheroid by combining number techniques, such as metabolic NADH Fluorescence Lifetime Imaging Microscopy (NADH-FLIM), hyperspectral solvatochromic lipophilic dye (Nile Red) matrix Second Harmonic Generation (SHG). included phasor-based bioimage analysis at three different time points, tracking both morphological biological properties, including cellular metabolism, fatty acids storage, collagen organization. Employing this deep-imaging framework, observed quantified cell plasticity response changes environment composition.

Language: Английский

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Comparison of vinculin tension in cellular monolayers and three-dimensional multicellular aggregates

Biomedical Optics Express, Journal Year: 2024, Volume and Issue: 15(9), P. 5199 - 5199

Published: Aug. 6, 2024

Confocal frequency-domain fluorescence lifetime and Förster resonance energy transfer (FRET) microscopy of Chinese hamster ovary (CHO-K1) cells expressing the vinculin tension sensor (VinTS) is used to compare in three-dimensional (3D) multicellular aggregates 2D cellular monolayers. In both 3D cultures, FRET efficiency VinTS 5-6% lower than that VinTL (p < 0.05), a tail-less control which cannot bind actin or paxillin. The difference between can be mitigated by treatment with Rho-associated kinase inhibitor Y-27632, demonstrating under cultures. However, there an overall decrease compared Expression cultures exhibits puncta consistent adhesions. While paxillin present at sites expression monolayers, it generally absent from aggregates. results suggest experiences modified environment monolayers provide basis for further investigation molecular sensors tissue models.

Language: Английский

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