Search Results (86)

Three-dimensional (3D) spheroid models have revolutionized in vitro cancer research by offering more physiologically relevant alternatives to traditional two-dimensional (2D) cultures. A systematic search identifies English-language studies on patient-derived cancer spheroids for drug screening, using defined inclusion and exclusion criteria, with data extracted on cancer type, culture methods, spheroid characteristics, and therapeutic responses. This manuscript evaluates the methods for spheroid formation and the cellular sources used, highlighting the diverse applications and preferences in this field. The five most investigated cancer origins for spheroid seeding are breast, colon, lung, ovary, and brain cancers, reflecting their clinical importance and research focus. Among seeding methodologies, forced-floating and scaffold-based methods predominate, demonstrating reliability and versatility in spheroid generation. Other techniques, including microfluidics, bioprinting, hanging drop, and suspension culture also play significant roles, each with distinct advantages and limitations. This review underscores the increasing use of spheroid models and the need for standardization in methodologies to enhance the reproducibility and translational potential in cancer research. Full article

The hanging-wall ramps of rift basins are prone to the accumulation of large sedimentary bodies and are potential areas for the presence of large subsurface geological reservoir volumes. This paper comprehensively utilizes data from sedimentology, seismic reflection, geochemistry, and palynology to study the paleotopography, water conditions, paleoclimate, and sediment supply of the fourth member (Mbr 4) of the Shahejie Formation in the Raoyang Sag of the Bohai Bay Basin, China. The sedimentary characteristics, evolution, and preserved stratigraphic architectures of shallow-water deltaic successions are analyzed. Multiple indicators—such as sporopollen, ostracoda, fossil algae, major elements, and trace elements—suggest that when Mbr 4 was deposited, the climate became progressively more humid, and the lake underwent deepening followed by shallowing. During rift expansion, the lake level began to rise with supplied sediment progressively filling available accommodation; sand delivery to the inner delta front was higher than in other parts of the delta, and highly active distributary channels formed a reticular drainage network on the delta plain, which was conducive to the formation of sandstone up-dip pinch-out traps. In the post-rift period, the lake water level dropped, and the rate and volume of sediment supply decreased, leading to the formation of a stable dendritic network of distributary channels. At channel mouths, sediments were easily reworked into sandsheets. The distribution of sandstone and mudstone volumes is characterized by up-dip pinch-out traps and sandstone lens traps. The network of channel body elements of the shallow-water deltaic successions is expected to act as an effective carbon dioxide storage reservoir. This study reveals the influence of multiple factors on the sedimentary characteristics, evolution, and internal network of shallow-water deltas at different stages of rift basin evolution. This knowledge helps improve resource utilization and the sustainable development of comparable subsurface successions. Full article

Spheroids, as three-dimensional (3D) cell aggregates, can be prepared using various methods, including hanging drops, microwells, microfluidics, magnetic manipulation, and bioreactors. However, current spheroid manufacturing techniques face challenges such as complex workflows, the need for specialized personnel, and poor batch reproducibility. In this study, we designed a support-free, 3D-printed microwell chip and developed a compatible low-cell-adhesion process. Through simulation and experimental validation, we rapidly optimized microwell size and the coating process. We successfully formed three types of spheroids—human immortalized epidermal cells (HaCaTs), umbilical cord mesenchymal stem cells (UC-MSCs), and human osteosarcoma cells (MG63s)—on the chip. Fluorescent viability staining confirmed the biocompatibility and reliability of the chip. Finally, drug response experiments were conducted using the chip. Compared to traditional methods, our proposed strategy enables high-throughput production of size-controlled spheroids with excellent shape retention, while enhanced gas exchange during culture improves differentiation marker expression. This platform provides an efficient and cost-effective solution for biosensing applications, such as drug screening, disease modeling, and personalized therapy monitoring. Furthermore, the chip shows significant potential for real-time in vitro monitoring of cellular viability, reaction kinetics, and drug sensitivity, offering valuable advancements in biosensor technology for life sciences and medical applications. Full article

Three-dimensional (3D) tissue culture models provide in vivo-like conditions for studying cell physiology. This study aimed to examine the efficiency of pyramidal microwell geometries in microfluidic devices on spheroid formation, cell growth, viability, and differentiation in mouse embryonic stem cells (mESCs). The static culture using the hanging drop (HD) method served as a control. The microfluidic chips were fabricated to have varying pyramidal tip angles, including 66°, 90°, and 106°. From flow simulations, when the tip angle increased, streamline distortion decreased, resulting in more uniform flow and a lower velocity gradient near the spheroids. These findings demonstrate the significant influence of microwell geometry on fluid dynamics. The 90° microwells provide optimal conditions, including uniform flow and reduced shear stress, while maintaining the ability for waste removal, resulting in superior spheroid growth compared to the HD method and other microwell designs. From the experiments, by Day 3, spheroids in the 90° microwells reached approximately 400 µm in diameter which was significantly larger than those in the 66° microwells, 106° microwells, and HD cultures. Brachyury gene expression in the 90° microwells was four times higher than the HD method, indicating enhanced mesodermal differentiation essential for cardiac differentiation. Immunofluorescence staining confirmed cardiomyocyte differentiation. In conclusion, microwell geometry significantly influences 3D cell culture outcomes. The pyramidal microwells with a 90° tip angle proved most effective in promoting spheroid growth and cardiac differentiation of mESC differentiation, providing insights for optimizing microfluidic systems in tissue engineering and regenerative medicine. Full article

Glioblastoma multiforme (GBM) is among the most aggressive brain cancers, and it contains glioma stem cells (GSCs) that drive tumor initiation, progression, and recurrence. These cells resist conventional therapies, contributing to high recurrence rates in GBM patients. Developing in vitro models that mimic the tumor microenvironment (TME), particularly the GSC niche, is crucial for understanding GBM growth and therapeutic resistance. Three-dimensional (3D) spheroid models provide a more physiologically relevant approach than traditional two-dimensional (2D) cultures, recapitulating key tumor features like hypoxia, cell heterogeneity, and drug resistance. This review examines scaffold-free and scaffold-based methods for generating 3D GBM spheroids, focusing on their applications in studying the cancer stem cell niche. The discussion encompasses methods such as the hanging drop, low-adhesion plates, and magnetic levitation, alongside advancements in embedding spheroids within extracellular matrix-based hydrogels and employing 3D bioprinting to fabricate more intricate tumor models. These 3D culture systems offer substantial potential for enhancing our understanding of GBM biology and devising more effective targeted therapies. Full article

Cell culture methods are indispensable strategies for studies in biological sciences and for drug discovery and testing. Most cell cultures have been developed using two-dimensional (2D) culture methods, but three-dimensional (3D) culture techniques enable the establishment of in vitro models that replicate various pathogenic conditions and they provide valuable insights into the pathophysiology of various diseases as well as more precise results in tests for drug efficacy. However, one difficulty in the use of 3D cultures is selection of the appropriate 3D cell culture technique for the study purpose among the various techniques ranging from the simplest single cell type-derived spheroid culture to the more sophisticated organoid cultures. In the simplest single cell type-derived spheroid cultures, there are also various scaffold-assisted methods such as hydrogel-assisted cultures, biofilm-assisted cultures, particle-assisted cultures, and magnet particle-assisted cultures, as well as non-assisted methods, such as static suspension cultures, floating cultures, and hanging drop cultures. Since each method can be differently influenced by various factors such as gravity force, buoyant force, centrifugal force, and magnetic force, in addition to non-physiological scaffolds, each method has its own advantages and disadvantages, and the methods have different suitable applications. We have been focusing on the use of a hanging drop culture method for modeling various non-cancerous and cancerous diseases because this technique is affected only by gravity force and buoyant force and is thus the simplest method among the various single cell type-derived spheroid culture methods. We have found that the biological natures of spheroids generated even by the simplest method of hanging drop cultures are completely different from those of 2D cultured cells. In this review, we focus on the biological aspects of single cell type-derived spheroid culture and its applications in in vitro models for various diseases. Full article

A hallmark of angiogenesis is the sprouting of endothelial cells. To replicate this event in vitro, biomaterial approaches can play an essential role in promoting cell migration. To study the capacity of a scaffold of fibrin (fibrinogen:thrombin mix) to support the movement of the endothelial cells, the migration area of spheroids formed with the HULEC cell line was measured. The cells were first allowed to form a spheroid using the hanging drop technique before being encapsulated in the fibrin gel. The cells’ migration area was then measured after two days of embedding in the fibrin gel. Various conditions affecting fibrin gel polymerization, such as different concentrations of fibrinogen and thrombin, were evaluated alongside rheology, porosity, and fiber thickness analysis to understand how these factors influenced cell behavior within the composite biomaterial. Data point toward thrombin’s role in governing fibrin gel polymerization; higher concentrations result in less rigid gels (loss tangent between 0.07 and 0.034) and increased cell migration (maximum concentration tested: 5 U/mL). The herein presented method allows for a more precise determination of the crosslinking conditions of fibrin gel that can be used to stimulate angiogenic sprouting. Full article

High-quality, 3D-shaped, SiO₂ colloidal photonic crystals (ellipsoids, hyperboloids, and others) were fabricated by self-assembly. They possess a quadratic surface and are wide-angle-independent, direction-dependent, diffractive reflection crystals. Their size varies between 1 and 5 mm and can be achieved as mechanical-resistant, free-standing, thick (hundreds of ordered layers) objects. High-quality, 3D-shaped, polystyrene inverse-opal photonic superstructures (highly similar to diatom frustules) were synthesized by using an inside infiltration method as wide-angle-independent, reflective diffraction objects. They possess multiple reflection bands given by their special architecture (a torus on the top of an ellipsoid) and by their different sized holes (384 nm and 264 nm). Our hanging-drop self-assembly approach uses setups which deform the shape of an ordinary spherical drop; thus, the colloidal self-assembly takes place on a non-axisymmetric liquid/air interface. The deformed drop surface is a kind of topological interface which changes its shape in time, remaining as a quality template for the self-assembly process. Three-dimensional-shaped colloidal photonic crystals might be used as devices for future spectrophotometers, aspheric or freeform diffracting mirrors, or metasurfaces for experiments regarding space-time curvature analogy. Full article

The European hazelnut (Corylus avellana L.) is a self-incompatible, wind-pollinated species of significant economic-productive interest, mainly cultivated between the Mediterranean basin and the Black Sea. Hazelnut breeding programs and high cropping in commercial orchards are necessarily linked to the availability of fertile pollen with wide germinability and high viability. The objective was to develop an improved method to determine the germinability of pollen, comparing the results with those found in the literature. Pollen germinability was firstly evaluated on a wild-type accession using boric acid (H₃BO₃), calcium nitrate (Ca(NO₃)₂), magnesium sulfate (MgSO₄), potassium nitrate (KNO₃), casein hydrolysate and different concentrations of sugars, including sucrose (10, 15 and 25%) and two monosaccharides, glucose (15%) and fructose (15%). The optimal composition included 15% sucrose in a semi-solid medium composed of 1% animal gelatin, containing collagen. This formulation was then tested on three cultivars of commercial interest, evaluating the effect of different concentrations of sucrose. The comparison between different in vitro germination methods showed that the hanging drop technique in a semi-solid medium provides a statistically higher germination index compared to the liquid and solid/agarized techniques. This study proposes an easy-to-use medium for the in vitro germination of Corylus avellana L. pollen. Full article

The Marangoni effect involves a mass transfer along an interface between two phases owing to the gradient of the interfacial tension. The flow caused by this phenomenon is called Marangoni convection, a complex phenomenon that involves mass transfer processes, such as surfactant adsorption/desorption processes, solvent dissolution phenomena, and viscous dissipation processes. Therefore, the strength of the convection depends on the various thermodynamic and physical properties of the fluids. In this study, we experimentally investigated the relationship between the Marangoni convection generated inside a hanging oil droplet and the interfacial tension of the oil droplet in an aqueous phase by the particle image velocimetry method. This convection velocity depended on the initial value of the interfacial tension in the oil–water interfacial tension oscillation phenomenon accompanied by the expansion and contraction of the hanging drop. Additionally, the droplet oscillation frequency decreased as the Marangoni convection velocity increased. Furthermore, continuous convection, which is unlike Marangoni convection, was observed within this spontaneously expanding and contracting hanging-droplet system. This buoyant convection was caused by the mutual dissolution of the hanging-droplet oil phase and the surrounding aqueous phase. Full article

Creating model systems that replicate in vivo tissues is crucial for understanding complex biological pathways like drug response and disease progression. Three-dimensional (3D) in vitro models, especially multicellular spheroids (MCSs), offer valuable insights into physiological processes. However, generating MCSs at scale with consistent properties and efficiently recovering them pose challenges. We introduce a workflow that automates large-scale spheroid production and enables parallel harvesting into individual wells of a microtiter plate. Our method, based on the hanging-drop technique, utilizes a non-contact dispenser for dispensing nanoliter droplets of a uniformly mixed-cell suspension. The setup allows for extended processing times of up to 45 min without compromising spheroid quality. As a proof of concept, we achieved a 99.3% spheroid generation efficiency and maintained highly consistent spheroid sizes, with a coefficient of variance below 8% for MCF7 spheroids. Our centrifugation-based drop transfer for spheroid harvesting achieved a sample recovery of 100%. We successfully transferred HT29 spheroids from hanging drops to individual wells preloaded with collagen matrices, where they continued to proliferate. This high-throughput workflow opens new possibilities for prolonged spheroid cultivation, advanced downstream assays, and increased hands-off time in complex 3D cell culture protocols. Full article

This pioneering study presents an in-depth biomechanical examinations of soccer’s diving header, aiming to quantify its impact on ball speed enhancement (BSE) and effective offensive range (EOR). Despite the diving header’s widespread acclaim and historical significance, there remains a dearth of scientific scrutiny into its biomechanical intricacies. Employing an innovative research design featuring a static hanging ball at varied offensive distances and heights, this study replicates diverse header scenarios. The results of 3D motion quantification have shown that a physically excellent player (identified through the maximal standing long jump test) could reach an EOR around 2.64 times his body height. Furthermore, this study unveils that proficient players could attain BSE surpassing 9 m/s, signifying the diving header’s heightened efficacy compared to traditional heading techniques, which could only result in 4.5 m/s. Correlation analyses unveil noteworthy relationships, highlighting the pivotal role of head speed at impact and the influence of minimizing speed drop and temporal disparities for amplified effectiveness. Considerations for optimizing diving header execution are introduced, emphasizing the necessity for targeted training programs. Despite acknowledged limitations inherent to its pilot nature, this exploration furnishes foundational knowledge to guide subsequent research and practical applications, providing valuable insights into soccer training and skill development through a biomechanical lens. Full article

High-quality convex colloidal photonic crystals can be grown on the tip of an optical fiber by self-assembly using the hanging drop method. They are convex-shaped, produce the diffraction of reflecting light with high efficiency (blazing colors), and have a high curvature. The convex colloidal crystals are easily detachable and, as free-standing objects, they are mechanically robust, allowing their manipulation and use as convex reflective diffraction devices in imaging spectrometers. Currently, the same characteristics are obtained by using gratings-based structures. The optical fiber/colloidal crystal interface is disordered; thus, no light diffraction can be registered. The ordering at this interface was highly increased by forming a polystyrene spacer on the optical fiber tip, which served as a self-assembly substrate for silica colloid, as a mechanical bond between the fiber and the crystal, and as a filler reservoir for an inverse-opal synthesis. The silica opal-like grown on the optical fiber tip can be transformed into a high-quality polystyrene (blazing colors) inverse-opal by using the polystyrene spacer as a filler. We found that the colloidal crystal axisymmetric self-assembles onto the optical fiber tip only if a maximum volume of the colloid drop is settled on a flat end of the polystyrene spacer. Full article

The encapsulation of stem cells into alginate microspheres is an important aspect of tissue engineering or bioprinting which ensures cell growth and development. We previously demonstrated the encapsulation of stem cells using the hanging drop method. However, this conventional process takes a relatively long time and only produces a small-volume droplet. Here, an experimental approach for alginate emulsification in multistage microfluidics is reported. By using the microfluidic method, the emulsification of alginate in oil can be manipulated by tuning the flow rate for both phases. Two-step droplet emulsification is conducted in a series of polycarbonate and polydimethylsiloxane microfluidic chips. Multistage emulsification of alginate for stem cell encapsulation has been successfully reported in this study under certain flow rates. Fundamental non-dimensional numbers such as Reynolds and capillary are used to evaluate the effect of flow rate on the emulsification process. Reynolds numbers of around 0.5–2.5 for alginate/water and 0.05–0.2 for oil phases were generated in the current study. The capillary number had a maximum value of 0.018 to ensure the formation of plug flow. By using the multistage emulsification system, the flow rates of each process can be tuned independently, offering a wider range of droplet sizes that can be produced. A final droplet size of 500–1000 µm can be produced using flow rates of 0.1–0.5 mL/h and 0.7–2.4 mL/h for the first stage and second stage, respectively. Full article

(1) Background: Acute aortic dissection (AAD) is caused by an endothelial entry tear followed by intimomedial delamination of the outer layers of the vessel wall. The established risk factors include hypertension and smoking. Another rising candidate risk factor is excessive alcohol consumption. This experimental study explores the effects of nicotine (Nic), angiotensin II (Ang II), and ethanol (EtOH) on human aortic endothelial cells (hAoEC). (2) Methods: HAoECs were exposed to Nic, Ang II, and EtOH at different dose levels. Cell migration was studied using the scratch assay and live-cell imaging. The metabolic viability and permeability capacity was investigated using the water-soluble tetrazolium (WST)-1 assay and an in vitro vascular permeability assay. Cell adherence was studied by utilizing the hanging drop assay. The transcriptional and protein level changes were analyzed by RT-qPCR, Western blotting and immunohistochemistry for major junctional complexing proteins. (3) Results: We observed reduced metabolic viability following Ang II and EtOH exposure vs. control. Further, cell adherence was enhanced by EtOH exposure prior to trituration and by all risk factors after trituration, which correlated with the increased gene and protein expression of VE-cadherin upon EtOH exposure. The cell migration capacity was reduced upon EtOH exposure vs. controls. (4) Conclusion: Marked functional changes were observed upon exposure to established and potential risk factors for AAD development in hAoECs. Our findings advocate for an enhanced mechanical rigidity in hAoECs in response to the three substances studied, which in turn might increase endothelial rigidity, suggesting a novel mechanism for developing an endothelial entry tear due to reduced deformability in response to increased shear and pulsatile stress. Full article