We are working on the development of functional test methods for cells and tissues, by application of our 3D culture and device technology. In particular, we provide values that cannot be obtained with conventional cellular testing methods, by means of higher-order functional tests that can quantitatively evaluate the functions of target cells and tissues (e.g., nerve transmission, muscle contraction, etc.). Through these methods, we aim to contribute to a wide range of basic and application fields, including basic research, drug discovery, regenerative medicine, and cosmetics. In addition to utilizing these technologies in our own drug discovery research, we are also accepting requests of joint research and contracted testing related to these technologies.
Schwann cells derived from human iPS cells
We conduct contract research and collaborative research using human iPS cell-derived Schwann cells established by our company. In addition to Schwann cells generated from healthy person-derived iPS cells, Schwann cells generated from Charcot-Marie-Tooth disease 1A (CMT1A) patient-derived iPS cells can also be used.
Feature
- Schwann cells showing high maturity
- High expression of MBP (Myelin Basic Protein)
- Produces NGF (Nerve Growth Factor)
Main applications
- Development and physiology of Schwann cells
- Co-culture with neurons
- Disease Modeling of CMT1A and Other Peripheral Neuropathy
- Pharmacological evaluation of compound efficacy for peripheral neuropathy and damage
Schwann cells derived from human iPS cell lines produced by our company
Multilayering of cells using oriented fiber scaffolds
By culturing cells on a scaffold of precisely oriented and controlled fiber sheets, it is possible to create multi-layered cell sheets in which cells are aligned along the fiber orientation. For example, when using nerve cells, a cell sheet with a thickness of 50~100 μm can be obtained with aligned nerve cells and axons stacked on the top and bottom of the fiber sheet. Such multi-layered neuronal cell sheets produced by our technology have superior features compared to 2D culture, such as (1) neural maturation is accelerated by the high-density culture of the 3D structure, and (2) stable adhesion to the scaffold inhibits cell aggregation and allows stable culture for longer assay periods with neural network activities.
(a) Orientation control by oriented fiber scaffolding
(b) Multilayering of nerve cells
Three-dimensional culture using oriented fiber sheets
Development of cellular devices
Our company develops and manufactures multiple tools and containers for cell and tissue culture for functional evaluation. For example, oriented fiber scaffolds and multilayered cell sheets as mentioned above were very thin and soft, making them unsuitable for handling with tweezers. In addition, some devices used for observation and measurement require specific measurement probes that are different from general culture vessels. In such cases, cell culture and measurement require specific probes adapted to each device, which raises issues of versatility and cost.
To solve these problems, we have developed a compact cell culture vessel that can be moved freely between various culture vessels and measurement devices. In addition, the container itself is structurally designed for easier evaluation of cell functions. For example, we developed the oriented fiber device, in which oriented fiber sheets are held in a small frame that fits into a 96-well plate. Such neural cell culture devices with oriented fiber sheets (SCAD devices) have been granted a U.S. patent right (patent number: US 11,543,404). We call them “cell devices” because they combine the functions of cell culture and functional evaluation and can be expanded to cell/tissue chips. Based on our microfabrication and plastic molding technologies, we have extended knowledge and experiences to carry out every step from material selection to design optimal shapes in-house, allowing us to flexibly design and produce prototypes according to the purpose and target.
(a) Example of a fiber device
(b) Cross-sectional view of a fiber device (image)
Fiber device developed by our company
Evaluation of electrophysiological properties using neuronal devices
Since the main function of neurons is to produce output and transmit electrical signals, it is important to measure normal or abnormal electrophysiological activities. One of such electrophysiology methods in recent years is measurement of extracellular potentials using a multi-electrode array (MEA). MEAs have microelectrodes arranged in an array and can simultaneously and continuously observe the extracellular potential of a cell population, making them suitable for evaluating neural network activity. However, when using neurons differentiated from iPS cells, the formation of neural networks requires a long culture period, and the desired results are often not obtained because the neurons detach from their containers and make aggregates during culture. Our multi-layered neuronal cell sheet fabricated using our 3D culture technology solves these problems and greatly enhances the practicality of the evaluation of electrophysiological properties of neural cell networks using MEA.
Utilization of SCAD neuronal devices for multi-electrode array measurement (early maturation by 3D culture)
The measurement of extracellular potential using MEA also gives a number of analytical parameters such as the number of spikes, synchronized burst firing frequency, synchronized burst duration, and so on. Since different analytical parameters are affected by different mechanisms of action of the test substance added to the cell culture, analysis of multidimensional data is required to determine the presence or absence of neurotoxicity and the therapeutic effect for neuropathy. We and our 3D cultured neural devices are capable of multidimensional analysis by principal component analysis and hierarchical clustering. Such usefulness of the measurement method using these neural devices was published in Biomaterial Research in 2022, co-authored with Professor Ikurou Suzuki of Tohoku Institute of Technology.
<External link>
「A functional neuron maturation device provides convenient application on microelectrode array for neural network measurement」
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