A magnetic levitation based low-gravity simulator with an unprecedented large functional volume
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ABSTRACT:
SUBMITTER: Sanavandi H
PROVIDER: S-EPMC8556250 | biostudies-literature |
REPOSITORIES: biostudies-literature
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Project description:Magnetic levitation, which uses a magnetic field to suspend objects in a fluid, is a powerful and versatile technology. We develop a compact magnetic levitation platform compatible with a smart-phone to separate micro-objects and estimate the density of the sample based on its levitation height. A 3D printed attachment is mechanically installed over the existing camera unit of a smart-phone. Micro-objects, which may be either spherical or irregular in shape, are suspended in a paramagnetic medium and loaded in a microcapillary tube which is then inserted between two permanent magnets. The micro-objects are levitated and confined in the microcapillary at an equilibrium height dependent on their volumetric mass densities (causing a buoyancy force toward the edge of the microcapillary) and magnetic susceptibilities (causing a magnetic force toward the center of the microcapillary) relative to the suspending medium. The smart-phone camera captures magnified images of the levitating micro-objects through an additional lens positioned between the sample and the camera lens cover. A custom-developed Android application then analyzes these images to determine the levitation height and estimate the density. Using this platform, we were able to separate microspheres with varying densities and calibrate their levitation heights to known densities to develop a technique for precise and accurate density estimation. We have also characterized the magnetic field, the optical imaging capabilities, and the thermal state over time of this platform.
| S-EPMC4550410 | biostudies-literature
Project description:Cell culture is an essential tool in drug discovery, tissue engineering and stem cell research. Conventional tissue culture produces two-dimensional cell growth with gene expression, signalling and morphology that can be different from those found in vivo, and this compromises its clinical relevance. Here, we report a three-dimensional tissue culture based on magnetic levitation of cells in the presence of a hydrogel consisting of gold, magnetic iron oxide nanoparticles and filamentous bacteriophage. By spatially controlling the magnetic field, the geometry of the cell mass can be manipulated, and multicellular clustering of different cell types in co-culture can be achieved. Magnetically levitated human glioblastoma cells showed similar protein expression profiles to those observed in human tumour xenografts. Taken together, these results indicate that levitated three-dimensional culture with magnetized phage-based hydrogels more closely recapitulates in vivo protein expression and may be more feasible for long-term multicellular studies.
| S-EPMC4487889 | biostudies-literature
Project description:Adulterated cooking oils and the repeated use of frying cooking oils are harmful to human health. Current techniques for the quality control of cooking oils such as gas chromatography and high performance liquid chromatography usually require expensive facilities, and they are complicated to operate. This paper describes a simple technique that uses magnetic levitation (MagLev) to analyse cooking oils based on density. We have demonstrated the application of MagLev to detect the quality of cooking oil by simulating adulterated sesame oil using peanut oil and frying soybean oil for different times. We have also demonstrated the use of MagLev to differentiate secondhand cooking oil from certified cooking oil. MagLev provides a portable and inexpensive method for the on-site inspection of cooking oils, and it may be extended to many other applications in food safety, environmental monitoring, medical diagnosis, and so on. The advantages of high sensitivity, low cost, and convenience of operation make MagLev especially useful for in situ applications in resource-limited settings.
| S-EPMC9064766 | biostudies-literature
Project description:BackgroundRecently, various studies have revealed that 3D cell spheroids have several advantages over 2D cells in stem cell culture. However, conventional 3D spheroid culture methods have some disadvantages and limitations such as time required for spheroid formation and complexity of the experimental process. Here, we used acoustic levitation as cell culture platform to overcome the limitation of conventional 3D culture methods.MethodsIn our anti-gravity bioreactor, continuous standing sonic waves created pressure field for 3D culture of human mesenchymal stem cells (hMSCs). hMSCs were trapped and aggerated in pressure field and consequently formed spheroids. The structure, viability, gene and protein expression of spheroids formed in the anti-gravity bioreactor were analyzed by electron microscope, immunostaining, polymerase chain reaction, and western blot. We injected hMSC spheroids fabricated by anti-gravity bioreactor into the mouse hindlimb ischemia model. Limb salvage was quantified to evaluate therapeutic efficacy of hMSC spheroids.ResultsThe acoustic levitation in anti-gravity bioreactor made spheroids faster and more compact compared to the conventional hanging drop method, which resulted in the upregulation of angiogenic paracrine factors of hMSCs, such as vascular endothelial growth factor and angiopoietin 2. Injected hMSCs spheroids cultured in the anti-gravity bioreactor exhibited improved therapeutic efficacy, including the degree of limb salvage, capillary formation, and attenuation of fibrosis and inflammation, for mouse hindlimb ischemia model compared to spheroids formed by the conventional hanging drop method.ConclusionOur stem cell culture system using acoustic levitation will be proposed as a new platform for the future 3D cell culture system.
| S-EPMC10197840 | biostudies-literature
Project description:Several cellular events cause permanent or transient changes in inherent magnetic and density properties of cells. Characterizing these changes in cell populations is crucial to understand cellular heterogeneity in cancer, immune response, infectious diseases, drug resistance, and evolution. Although magnetic levitation has previously been used for macroscale objects, its use in life sciences has been hindered by the inability to levitate microscale objects and by the toxicity of metal salts previously applied for levitation. Here, we use magnetic levitation principles for biological characterization and monitoring of cells and cellular events. We demonstrate that each cell type (i.e., cancer, blood, bacteria, and yeast) has a characteristic levitation profile, which we distinguish at an unprecedented resolution of 1 × 10(-4) g ? mL(-1). We have identified unique differences in levitation and density blueprints between breast, esophageal, colorectal, and nonsmall cell lung cancer cell lines, as well as heterogeneity within these seemingly homogenous cell populations. Furthermore, we demonstrate that changes in cellular density and levitation profiles can be monitored in real time at single-cell resolution, allowing quantification of heterogeneous temporal responses of each cell to environmental stressors. These data establish density as a powerful biomarker for investigating living systems and their responses. Thereby, our method enables rapid, density-based imaging and profiling of single cells with intriguing applications, such as label-free identification and monitoring of heterogeneous biological changes under various physiological conditions, including antibiotic or cancer treatment in personalized medicine.
| S-EPMC4507238 | biostudies-literature
Project description:Multicellular spheroids are obtained in a variety of three-dimensional (3D) culture systems without the use of supporting scaffold. We present here a 3D culture method that resulted in a multicellular sheet under scaffold-free conditions. A floating disk-shaped 3D culture was prepared by magnetic levitation of B16F10 cells that has ingested Fe3O4-containing fibroin microspheres. The melanoma disk grew up to 19 mm in diameter and the thickness was ranged between 80 and 100 ?m. The 3D culture was filled with closely packed cells that were proliferating exponentially at a specific growth rate of µ = 0.015 h-1. Approximately half of the cells were Ki-67 positive with no detectable levels of apoptotic or autophagic cells. However, the percentage of propidium iodide-permeable cells was 8.5 ± 1.2 %, which was probably due to physical damage in the cell membrane caused by Fe3O4-containing microspheres under a strong magnetic field. Melanin production increased by a factor of 3.0-3.7 in the 3D culture, due to an increased population of pigmented cells. This study presented a surface 3D culture of B16F10 cells without the use of a scaffold based on magnetic levitation.
| S-EPMC5101302 | biostudies-literature
Project description:White adipose tissue (WAT) is becoming widely used in regenerative medicine/cell therapy applications, and its physiological and pathological importance is increasingly appreciated. WAT is a complex organ composed of differentiated adipocytes, stromal mesenchymal progenitors known as adipose stromal cells (ASC), as well as endothelial vascular cells and infiltrating leukocytes. Two-dimensional (2D) culture that has been typically used for studying adipose cells does not adequately recapitulate WAT complexity. Improved methods for reconstruction of functional WAT ex vivo are instrumental for understanding of physiological interactions between the composing cell populations. Here, we used a three-dimensional (3D) levitation tissue culture system based on magnetic nanoparticle assembly to model WAT development and growth in organoids termed adipospheres. We show that 3T3-L1 preadipocytes remain viable in spheroids for a long period of time, while in 2D culture, they lose adherence and die after reaching confluence. Upon adipogenesis induction in 3T3-L1 adipospheres, cells efficiently formed large lipid droplets typical of white adipocytes in vivo, while only smaller lipid droplet formation is achievable in 2D. Adiposphere-based coculture of 3T3-L1 preadipocytes with murine endothelial bEND.3 cells led to a vascular-like network assembly concomitantly with lipogenesis in perivascular cells. Adipocyte-depleted stromal vascular fraction (SVF) of mouse WAT cultured in 3D underwent assembly into organoids with vascular-like structures containing luminal endothelial and perivascular stromal cell layers. Adipospheres made from primary WAT cells displayed robust proliferation and complex hierarchical organization reflected by a matricellular gradient incorporating ASC, endothelial cells, and leukocytes, while ASC quickly outgrew other cell types in adherent culture. Upon adipogenesis induction, adipospheres derived from the SVF displayed more efficient lipid droplet accumulation than 2D cultures. This indicates that 3D intercellular signaling better recapitulates WAT organogenesis. Combined, our studies show that adipospheres are appropriate for WAT modeling ex vivo and provide a new platform for functional screens to identify molecules bioactive toward individual adipose cell populations. This 3D methodology could be adopted for WAT transplantation applications and aid approaches to WAT-based cell therapy.
| S-EPMC3603558 | biostudies-literature
Project description:Magnetic levitation (MagLev) has recently emerged as a powerful method to develop diagnostic technologies based on the exploitation of the nanoparticle (NP)-protein corona. However, experimental procedures improving the robustness, reproducibility, and accuracy of this technology are largely unexplored. To contribute to filling this gap, here, we investigated the effect of total flow rate (TFR) and flow rate ratio (FRR) on the MagLev patterns of microfluidic-generated graphene oxide (GO)-protein complexes using bulk mixing of GO and human plasma (HP) as a reference. Levitating and precipitating fractions of GO-HP samples were characterized in terms of atomic force microscopy (AFM), bicinchoninic acid assay (BCA), and one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (1D SDS-PAGE), and nanoliquid chromatography-tandem mass spectrometry (nano-LC-MS/MS). We identified combinations of TFR and FRR (e.g., TFR = 35 μL/min and FRR (GO:HP) = 9:1 or TFR = 3.5 μL/min and FRR (GO:HP) = 19:1), leading to MagLev patterns dominated by levitating and precipitating fractions with bulk-like features. Since a typical MagLev experiment for disease detection is based on a sequence of optimization, exploration, and validation steps, this implies that the optimization (e.g., searching for optimal NP:HP ratios) and exploration (e.g., searching for MagLev signatures) steps can be performed using samples generated by bulk mixing. When these steps are completed, the validation step, which involves using human specimens that are often available in limited amounts, can be made by highly reproducible microfluidic mixing without any ex novo optimization process. The relevance of developing diagnostic technologies based on MagLev of coronated nanomaterials is also discussed.
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Project description:Chronic infections are associated with the formation of nonattached biofilm-like aggregates. In vitro models of surface-attached biofilms do not always accurately mimic these processes. Here, we tested a new approach to create in vitro nonattached bacterial aggregates using the principle of magnetic levitation of biological objects placed into a magnetic field gradient. Bacteria grown under magnetic levitation conditions formed nonattached aggregates that were studied with confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM) and characterized quantitatively. Nonattached aggregates consisted of bacteria submerged into an extracellular matrix and demonstrated features characteristic of biofilms, such as a polymeric matrix that binds Ruby Red and Congo red dyes, a prerequisite of bacterial growth, and increased resistance to gentamicin. Three quantitative parameters were explored to characterize strain-specific potential to form nonattached aggregates: geometric sizes, relative quantities of aggregated and free-swimming bacteria, and Congo red binding. Among three tested Escherichia coli strains, one strain formed nonattached aggregates poorly, and for this strain, all three of the considered parameters were different from those of the other two strains (P?<?0.05). Further, we characterized biofilm formation on plastic and agar surfaces by these strains and found that good biofilm formation ability does not necessarily indicate good nonattached aggregate formation ability, and vice versa. The model and quantitative methods can be applied for in vitro studies of nonattached aggregates and modeling bacterial behavior in chronic infections, as it is important to increase our understanding of the role that nonattached bacterial aggregates play in the pathogenesis of chronic diseases.IMPORTANCE An increasing amount of evidence indicates that chronic infections are associated with nonattached biofilm-like aggregates formed by pathogenic bacteria. These aggregates differ from biofilms because they form under low-shear conditions within the volume of biological fluids and they do not attach to surfaces. Here, we describe an in vitro model that provides nonattached aggregate formation within the liquid volume due to magnetic levitation. Using this model, we demonstrated that despite morphological and functional similarities of nonattached aggregates and biofilms, strains that exhibit good biofilm formation might exhibit poor nonattached aggregate formation, suggesting that mechanisms underlying the formation of biofilms and nonattached aggregates are not identical. The magnetic levitation approach can be useful for in vitro studies of nonattached aggregate formation and simulation of bacterial behavior in chronic infections.
| S-EPMC7480373 | biostudies-literature
Project description:Description Miniaturized and soft magnetically levitated centrifugal pumps can handle various fluids for wearable applications. Centrifugal pumps are essential mechanical components for liquid delivery in many biomedical systems whose miniaturization can promote innovative disease treatment approaches. However, centrifugal pumps are predominately constructed by rigid and bulky components. Here, we combine the soft materials and flexible electronics to achieve soft magnetic levitation micropumps (SMLMs) that are only 1.9 to 12.8 grams in weight. The SMLMs that rotate at a rotation speed of 1000 revolutions per min to pump liquids with various viscosities ranging from 1 to 6 centipoise can be used in assisting dialysis, blood circulation, and skin temperature control because of excellent biocompatibility with no organ damage. The development of SMLMs not only demonstrates the possibility to replace rigid rotating structures with soft materials for handling large volumes of fluids but also indicates the potential for fully flexible artificial organs that may revolutionize health care and improve the well-being of patients.
| S-EPMC8550243 | biostudies-literature