Yuya Morimoto

Molding Cell Beads for Rapid Construction of Macroscopic 3D Tissue Architecture

A microfluidic system was used to prepare a large number of size‐controlled collagen gel beads to form microtissue units, “cell beads”, as tissue building blocks. By stacking cell beads into a doll‐shaped silicone chamber, millimeter‐thick tissue with uniform cell dens...

Monodisperse cell-encapsulating peptide microgel beads for 3D cell culture.

This paper describes a method to produce monodisperse cell-encapsulating microgel beads composed of a self-assembling peptide gel for three-dimensional (3D) cell culture. We used a 3D microfluidic axisymmetric flow-focusing device with an external gelation method. The finely powdered salts were dispersed into a continuous phase, and the salts induced the gelation when in contact with the peptide solution. Over 93% of the cells survived after the encapsulation, and the cells migrated and grew within the gels. Applications of our cell-encapsulating beads include bead-based cell assays in drug testing and engineering tissue constructs.

Three-dimensional neuron–muscle constructs with neuromuscular junctions

This paper describes a fabrication method of muscle tissue constructs driven by neurotransmitters released from activated motor neurons. The constructs consist of three-dimensional (3D) free-standing skeletal muscle fibers co-cultured with motor neurons. We differentiated mouse neural stem cells (mNSCs) cultured on the skeletal muscle fibers into neurons that extend their processes into the muscle fibers. We found that acetylcholine receptors (AChRs) were formed at the connection between the muscle fibers and the neurons. The neuron-muscle constructs consist of highly aligned, long and matured muscle fibers that facilitate wide contractions of muscle fibers in a single direction. The contractions of the neuron-muscle construct were observed after glutamic acid activation of the neurons. The contraction was stopped by treatment with curare, an neuromuscular junction (NMJ) antagonist. These results indicate that our method succeeded in the formation of NMJs in the neuron-muscle constructs. The neuron-muscle construct system can potentially be used in pharmacokinetic assays related to NMJ disease therapies and in soft-robotic actuators.

Millimeter-Sized Neural Building Blocks for 3D Heterogeneous Neural Network Assembly

A millimeter-sized neural building block (NBB) shows high versatility to form a 3D heterogeneous neural component. A millimeter-sized 3D neural network between heterogeneous neural tissues is established, and an efficient technique is then developed to observe the spatiotemporal metrological changes of single neuron in the NBB. This technique allows the visualization of axonal extension, dendritic branching, and morphological changes of presynaptic components and synapses in real time.

Three-dimensional cell culture based on microfluidic techniques to mimic living tissues

This mini-review consists of microfluidic fabrication methods of cellular spheroids and cell-laden hydrogels, and their applications for tissue engineering. Using microfluidic devices, cellular spheroids and cell-laden hydrogels with controllable design are formed reproducibly. Owing to their size uniformity, they are used as building blocks for bottom-up tissue engineering to construct uniform and arbitrarily shaped tissues. Thus, cellular spheroids and cell-laden hydrogels based on microfluidic techniques are powerful tools to create tissues for human implantation and the treatment of diseases.

Skin integrated with perfusable vascular channels on a chip.

This paper describes a method for fabricating perfusable vascular channels coated with endothelial cells within a cultured skin-equivalent by fixing it to a culture device connected to an external pump and tubes. A histological analysis showed that vascular channels were constructed in the skin-equivalent, which showed a conventional dermal/epidermal morphology, and the endothelial cells formed tight junctions on the vascular channel wall. The barrier function of the skin-equivalent was also confirmed. Cell distribution analysis indicated that the vascular channels supplied nutrition to the skin-equivalent. Moreover, the feasibility of a skin-equivalent containing vascular channels as a model for studying vascular absorption was demonstrated by measuring test molecule permeation from the epidermal layer into the vascular channels. The results suggested that this skin-equivalent can be used for skin-on-a-chip applications including drug development, cosmetics testing, and studying skin biology.

Construction of 3D, Layered Skin, Microsized Tissues by Using Cell Beads for Cellular Function Analysis

Microsized skin cell beads consisting of 3D layered structures with epidermal and dermal cells have been fabricated. The beads are monodisperse, type-I collagen beads encapsulating dermal cells and covered by epidermal cells. The beads can be used in individual analyses of cellular functions and respond to chemical stimulation.

Point-, line-, and plane-shaped cellular constructs for 3D tissue assembly.

Microsized cellular constructs such as cellular aggregates and cell-laden hydrogel blocks are attractive cellular building blocks to reconstruct 3D macroscopic tissues with spatially ordered cells in bottom-up tissue engineering. In this regard, microfluidic techniques are remarkable methods to form microsized cellular constructs with high production rate and control of their shapes such as point, line, and plane. The fundamental shapes of the cellular constructs allow for the fabrication of larger arbitrary-shaped tissues by assembling them. This review introduces microfluidic formation methods of microsized cellular constructs and manipulation techniques to assemble them with control of their arrangements. Additionally, we show applications of the cellular constructs to biological studies and clinical treatments and discuss future trends as their potential applications.

A hybrid axisymmetric flow-focusing device for monodisperse picoliter droplets

A combination of photolithography and stereolithography was successfully used to fabricate a hybrid axisymmetric flow-focusing device (h-AFFD) that produces monodisperse picoliter droplets. The h-AFFD achieved the same level of hydrodynamic performance as a monolithic AFFD produced by only stereolithography from acrylic resin. Since the h-AFFD had a narrower orifice (50 or 100 ?m in diameter), created in an SU-8 sheet by photolithography, than the monolithic AFFD, we were able to produce picoliter droplets. We also succeeded in producing monodisperse droplets encapsulating a single cell without any surface modification.

Liquid-filled tunable lenticular lens

This paper describes a liquid-filled tunable lenticular lens for switching between two-dimensional (2D) and three-dimensional (3D) images in naked-eye 3D displays. Compared with previous 2D/3D switchable displays, this tunable lenticular lens that is directly attached to a smartphone display can project both a 2D image with the original resolution of the smartphone display and a 3D image with high brightness. This lens is simply composed of transparent poly(dimethylsiloxane) (PDMS) microchannels. While the thin top membrane on the microchannels is normally flat to transmit light without deflection for displaying 2D images, applying pressure to the microchannel deforms the membrane to acquire characteristics of lenticular lenses for 3D images. We successfully demonstrate the switching between the 2D and 3D modes. We believe that our lens can be applied as a part of a portable 2D/3D naked-eye 3D display.