A multiparametric analysis including single-cell and subcellular feature assessment reveals differential behavior of spheroid cultures on distinct ultra-low attachment plate types DOI Creative Commons
Mario Vitacolonna, Roman Bruch,

Ane Agaçi

et al.

Frontiers in Bioengineering and Biotechnology, Journal Year: 2024, Volume and Issue: 12

Published: Aug. 2, 2024

Spheroids have become principal three-dimensional models to study cancer, developmental processes, and drug efficacy. Single-cell analysis techniques emerged as ideal tools gauge the complexity of cellular responses in these models. However, single-cell quantitative assessment based on 3D-microscopic data subcellular distribution fluorescence markers, such nuclear/cytoplasm ratio transcription factors, has largely remained elusive. For spheroid generation, ultra-low attachment plates are noteworthy due their simplicity, compatibility with automation, experimental commercial accessibility. it is unknown whether what degree plate type impacts formation biology. This developed a novel AI-based pipeline for 3D-confocal optically cleared large spheroids at wholemount, single-cell, sub-cellular levels. To identify relevant samples pipeline, automated brightfield microscopy was employed systematically compare size eccentricity formed six different types using four distinct human cell lines. showed that all exhibited similar spheroid-forming capabilities gross patterns growth or shrinkage during 4 days after seeding were comparable. Yet, varied among specific lines types. Based this prescreen, HaCaT keratinocytes HT-29 cancer cells further assessed. In spheroids, in-depth revealed correlation between size, proliferation, transcriptional coactivator, YAP1, well an inverse respect differentiation. These findings, yielded model level, corroborate earlier concepts role YAP1 proliferation differentiation skin. Further, results show may influence outcome campaigns advisable scan optimal configuration investigation.

Language: Английский

Spheroids in cancer research: Recent advances and opportunities DOI Creative Commons
Sanchit Arora,

Somay Singh,

Anuj Mittal

et al.

Journal of Drug Delivery Science and Technology, Journal Year: 2024, Volume and Issue: 100, P. 106033 - 106033

Published: Aug. 10, 2024

In recent years, three-dimensional (3D) spheroid technology has emerged as a crucial instrument in the realm of cancer research, offering near-physiological model for probing human cancers. By faithfully replicating organ architecture and functionality, spheroids furnish versatile platform addressing spectrum clinical biomedical inquiries, encompassing pharmacology disease pathophysiology. Their distinct advantage over conventional two-dimensional (2D) cell cultures lies their capacity to emulate 3D extracellular microenvironment attributes characteristic solid tumors, including architectural intricacies, gene expression profiles, secretion soluble mediators. Derived effectively from both normal malignant patient tissues, facilitate modeling progression, mutation dynamics, carcinogenesis pathways. Moreover, expedites drug screening processes personalized therapeutic interventions. Although challenges persist accurately recapitulating immune system within models, co-culturing with lymphocytes holds significant promise immunotherapy applications. This comprehensive examination outlines diverse methodologies establishing characterizing spheroids, highlighting extensive utilization oncology. The manuscript underscores immense paving way an uprising understanding management cancer, abundant opportunities further investigation progress treatment approaches.

Language: Английский

Citations

8
Advancement in 3D Printable Materials for the Management of Cancer: A New Era of Materialistic Approach for the Treatment of Cancer DOI

Rishav Sharma,

S. Logesh Kumar, Rishabha Malviya

et al.

Journal of Drug Delivery Science and Technology, Journal Year: 2024, Volume and Issue: 100, P. 106064 - 106064

Published: Aug. 14, 2024

Language: Английский

Citations

6
Bridging high resolution sub-cellular imaging with physiologically relevant engineered tissues DOI

Yasaman Kargar Gaz Kooh,

Nathaniel Huebsch

SSRN Electronic Journal, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 1, 2025

While high-resolution microscopic techniques are crucial for studying cellular structures in cell biology, obtaining such images from thick 3D engineered tissues remains challenging. In this review, we explore advancements fluorescence microscopy, alongside the use of various fluorescent probes and material processing to address these challenges. We navigate through diverse array imaging options available tissue engineering field, wide field super-resolution so researchers can make more informed decisions based on specific interest. Finally, provide some recent examples how traditional limitations sub-cellular architecture within have been overcome by combining with innovative approaches.

Language: Английский

Citations

0
In vitro co-culture of Fasciola hepatica newly excysted juveniles (NEJs) with 3D HepG2 spheroids permits novel investigation of host–parasite interactions DOI Creative Commons
Aiste Vitkauskaite, Emma McDermott, Richard Lalor

et al.

Virulence, Journal Year: 2025, Volume and Issue: 16(1)

Published: March 25, 2025

Fasciola hepatica, or liver fluke, causes fasciolosis in humans and livestock. Following ingestion of vegetation contaminated with encysted parasites, metacercariae, newly excysted juveniles (NEJ) excyst the small intestine cross intestinal wall. After penetrating liver, parasite begins an intra-parenchymal migratory feeding phase that not only drives their rapid growth development but also extensive haemorrhaging immune pathology. Studies on infection are hindered by difficulty accessing these microscopic juvenile parasites vivo. Thus, a simple scalable vitro culture system for is needed. Here, we find two-dimensional (2D) systems using cell monolayers support NEJ to limited extent. By contrast, co-culture F. hepatica HepG2-derived 3D spheroids, "mini-livers," more closely mimic physiology microenvironment vivo tissue, promoted survival, growth, development. grazed peripheral cells they released temporally regulated digestive cysteine proteases, FhCL3, FhCL1/2, similar parasites. The induced gut body musculature, stimulated tegument elaborate spines variety surface sensory/tango/chemoreceptor papillae (termed S1, S2, S3); were especially pronounced around oral ventral suckers sense host chemical cues secure tissue. HepG2 spheroid/parasite methodologies should accelerate investigations into understanding developmental biology studies host–parasite interactions, streamline search new anti-parasite interventions.

Language: Английский

Citations

0
Bridging high resolution sub-cellular imaging with physiologically relevant engineered tissues DOI Creative Commons

Yasaman Kargar Gaz Kooh,

Nathaniel Huebsch

Frontiers in Mechanical Engineering, Journal Year: 2024, Volume and Issue: 10

Published: Nov. 26, 2024

While high-resolution microscopic techniques are crucial for studying cellular structures in cell biology, obtaining such images from thick 3D engineered tissues remains challenging. In this review, we explore advancements fluorescence microscopy, alongside the use of various fluorescent probes and material processing to address these challenges. We navigate through diverse array imaging options available tissue engineering field, wide field super-resolution so researchers can make more informed decisions based on specific interest. Finally, provide some recent examples how traditional limitations sub-cellular architecture within have been overcome by combining with innovative approaches.

Language: Английский

Citations

1
Experimental modeling, prevention, and treatment of endometriosis-associated adhesion formation in NIH/3T3 fibroblast cell line using bovhyaluronidase azoximer (in vitro) DOI
L.V. Adamyan, L.G. Pivazyan, K.S. Mailova

et al.

Russian Journal of Human Reproduction, Journal Year: 2024, Volume and Issue: 30(6), P. 61 - 61

Published: Jan. 1, 2024

Language: Английский

Citations

1
A multiparametric analysis including single-cell and subcellular feature assessment reveals differential behavior of spheroid cultures on distinct ultra-low attachment plate types DOI Creative Commons
Mario Vitacolonna, Roman Bruch,

Ane Agaçi

et al.

Frontiers in Bioengineering and Biotechnology, Journal Year: 2024, Volume and Issue: 12

Published: Aug. 2, 2024

Spheroids have become principal three-dimensional models to study cancer, developmental processes, and drug efficacy. Single-cell analysis techniques emerged as ideal tools gauge the complexity of cellular responses in these models. However, single-cell quantitative assessment based on 3D-microscopic data subcellular distribution fluorescence markers, such nuclear/cytoplasm ratio transcription factors, has largely remained elusive. For spheroid generation, ultra-low attachment plates are noteworthy due their simplicity, compatibility with automation, experimental commercial accessibility. it is unknown whether what degree plate type impacts formation biology. This developed a novel AI-based pipeline for 3D-confocal optically cleared large spheroids at wholemount, single-cell, sub-cellular levels. To identify relevant samples pipeline, automated brightfield microscopy was employed systematically compare size eccentricity formed six different types using four distinct human cell lines. showed that all exhibited similar spheroid-forming capabilities gross patterns growth or shrinkage during 4 days after seeding were comparable. Yet, varied among specific lines types. Based this prescreen, HaCaT keratinocytes HT-29 cancer cells further assessed. In spheroids, in-depth revealed correlation between size, proliferation, transcriptional coactivator, YAP1, well an inverse respect differentiation. These findings, yielded model level, corroborate earlier concepts role YAP1 proliferation differentiation skin. Further, results show may influence outcome campaigns advisable scan optimal configuration investigation.

Language: Английский

Citations

0