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This paper describes a method of sequential formation of asymmetric lipid vesicles, which applies pulse jet flow to an asymmetric planar lipid bilayer with multiple lipid components formed by Droplet Split-and-Contact Method. In our previous work, we generated asymmetric lipid vesicles by deforming an asymmetric planar lipid bilayer using a pulsed jet flow. However, it was difficult to create various...
In this paper, we developed a centrifugal emulsification device toward high-throughput generation of monodisperse liposomes. The device generated lipid vesicles in two steps, membrane emulsification by asymmetric through-holes on a plate, and transfer of the emulsions to a planar monolayer at an oil/water interface. As a result of centrifugation of the device in a microtube, we succeeded in collection...
This research reports a method to fabricate a loop-shaped vessel-like channel for perfusion of culture medium in a cell-laden collagen gel. We used alginate fibers as sacrificial structures for construction of loop-shaped vessel-like channels. By integration of a loop-shaped alginate fiber with connection ports in a cell-laden collagen gel, we formed a tube-connectable loop-shaped channel. Finally,...
We propose a flexible glucose sensor combined with a parylene based electrode and a glucose-responsive fluorescent hydrogel for continuous monitoring of glucose concentration. By depositing parylene-C on a cupper (Cu) thin sheet and creating electrical circuits including a LED and a photo diode, we developed the flexible glucose sensor. The flexible glucose sensor could reduce damage to tissues and...
This work presents a method to detect volatile odorant using corneal epithelial cells with olfactory receptors. Because the corneal epithelial cells can survive at constant exposure of air, these cells have ability to detect the volatile odorant at air-liquid phase in contrast to other parenchymal cells. Therefore, in the present study, we first fabricate a device that provides the air-liquid environment...
A rapid and high-resolution bioprinter based on a capillary coaxial microfluidic printer head and vacuum substrate, was conceived and built up in this work. We demonstrate the printing of porous (400 μm pitch) 3D bio-scaffolds made from calcium alginate microfibers with a printing resolution (i.e. fiber diameter) of ca. 150 microns, at a speed of 40 mm/s, which is approximately 10x faster than existing...
This paper reports on a biomimetic microskeleton actuated by muscle fibers (figure 1(a)). The 3D microskeleton is based on concatenated rib elements fabricated by two-photon polymerization of a custom-made hydrogel (figure 1(b)). For the first time, micrometer-sized skeletons were fabricated, overgrown with functional muscle fibers, and contracted at significant amplitudes. Compared to planar biomimetic...
This paper describes a core-shell hydrogel microfiber consisted of alginate and tetra-arm poly(ethylene glycol) (tetra-PEG) encapsulating rat insulinoma cells using a microfluidic process with a coaxial device and rapid gelation induced by diffusion of two different cross-linkers of barium ion and dithiothreitol (DTT) in the laminar flow (Fig. 1). Microfluidic techniques are able to encapsulate the...
This study reports an efficient droplet interface bilayer (DIB) formation using an open chamber with an external separator that can divide an aqueous solution to two regions. This approach facilitates reagent-saving and simplification of the device fabrication. Amphiphobic modification of the separator surface enables us to handle the device easily without liquid spill, and assists bilayer lipid membrane...
We developed the glucose-responsive florescent tetra-PEG gel based on diboronic acid for implantable continuous glucose monitoring. The fluorescent dye having diamine unit was immobilized into tetra-PEG gel formed by mixture of tetraamine-terminated-PEG (TAPEG) and tetra-NHS-glutarate-terminated-PEG (TNPEG). When the glucose concentration increased from 0 mg•dL−1 to 1,000 mg•dL−1, the fluorescence...
We developed a portable odorant detection system composed of collagen micro-pillars with cells expressing olfactory receptors and a CMOS imaging sensor; the CMOS imaging sensor caught fluorescence signals of Ca2+ increased by odorant molecules. While low sensitiveness of the CMOS imaging sensor was insufficient to detect the fluorescent signal from a single cell, the odorant detection system achieved...
MicroRNA, a short-length RNA known as a biomarker of cancers, was detected by magnet-attached spiral channel and electrical measurement. A designed oligo-DNA bound with magnetic bead was entrapped in spiral channel by magnetic field, and hybridized with microRNA. Thereafter, liposome were exported from the spiral channel by duplex-specific nuclease, and applied to electrical measurement. Finally,...
This paper highlights the behavior of microdroplets used for portable biosensors. Microdroplets have been applied for various Lab-on-a-chip applications, in which portability/wearable is becoming a trend word. However, the resonance (sloshing phenomena) of the microdroplets could be an issue. For example, bilayer lipid membrane, fragile characteristics due to its thickness of 5 nm, easily ruptures...
This article confirmed that the sensor retains its measurement capability for more than 150 days in a saline solution. In addition, an optimization of the sensor structure for obtaining an improved response time was carried out. A response time as short as that of the conventional self-monitoring of blood glucose (SMBG) technology was obtained with the optimized sensor structure.
We propose a drug testing system for quantifying contractile frequency and force of fiber-shaped tissues of human iPS-derived cardiomyocytes in various drugs. By optimization of cell culture conditions for the cardiomyocytes fiber and dimensions of the PDMS devices with cantilevers, we succeeded in measurement of contractile frequency and force derived from deformations of the cantilevers. As a result...
This paper describes a preliminary result regarding formation of core-shell microparticles utilizing a centrifuge-based microfluidic device. As a proof of concept, we encapsulated and cultured human dermal fibroblast (HDF) cells into such microstructures. In order to fabricate multi-compartmental microparticles1, the easy microparticles centrifugal synthesis method, which requires a tabletop centrifuge...
This paper presents a new method to align collagen nanofibers on arbitrary-shaped 3-dimensional (3D) surfaces, and applies the method to align collagen nanofibers and cells in circumferential directions on cylindrical surfaces. In this work, collagen nanofibers are self-aligned on 3D surfaces by applying cyclic stretch during the gelation process of collagen solution. Furthermore, the aligned collagen...
This paper reports on a 3D bio-hybrid metamaterial MEMS scaffold for the culture and differentiation of C2C12 muscle precursor cells. For the first time, preferential cells differentiation on a technical surface into functional myotubes is demonstrated. The micro-scaffolds are based on concatenated bowtie elements fabricated by two-photon lithography and designed to achieve an equivalent Young's modulus...
This paper proposes a self-assembly method to construct a droplet interface bilayer (DIB) network. The DIB network has been developed for biological logical circuits or chemical microreactors, using incorporated membrane proteins. In this work, the DIB network was constructed by using multi-geometric Droplet-Boxes (DBs), which captured and stabilized an aqueous droplet in organic solvent. The multi-geometry...
It is expected that induced pluripotent stem(iPS) cells contribute regenerative medicine. To reconstruct organ-like structure and function in vitro, the technological development of formation of three dimensional(3D) iPS cell culture and differentiation system has been required. Recently core-shell hydrogel microfiber system was reported in which cells were cultured with forming fibrous structure...
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