The team is working on generating alveolar organoids from isolated mouse alveolar stem cells, as well as tracheal mesenchymal organoids from ES and iPS cells. Using alveolar organoids, they are carrying out research on stemness and heterogeneity of tissue stem cells, as well as research of disease models. With tracheal mesenchyme organoids, they are developing 3D culture methods for inducing cartilage tissue structures and aiming to understand mechanisms of tracheal cartilage patterning.

This team is studying self-organization mechanisms during nephron formation by quantitatively analyzing organ deformation and individual cell dynamics based on 4D imaging data of kidney organoid formation from human iPS cells and of mouse embryonic kidney development. In particular, we aim to provide clues for realizing organoid maturation by clarifying how the dynamics during the formation of organoid whose shape/size is immature differs from that during normal development. This study is performed in collaboration with Takasato Team.

Tubules are fundamental units of our bodies and all metazoans. Its formation in organoid cultures is still imperfect in general. Axial elongation and uniform inner diameter are two geometrical characteristics of tubules. While tubulation and elongation are known to be controlled by external cues, how inner diameter is regulated is poorly understood. In order to gain deep understanding of tubule formation mechanisms, we use tracheal tubulogenesis in Drosophila as a model system. We address the structural basis of materials inside of the tube lumen and mechanical cross talk between luminal matrix and tubule epithelial cells to establish tube geometry.

The team is working to establish a new approach for self-organization-based induction of primordial organs and organoids, referred to as ‘direct reprogramming organogenesis.’ In addition, they are also striving to understand the mechanisms underlying organogenesis, particularly the molecular mechanisms inducing periodic morphological changes of hair follicles. They aim to cultivate primordial organs and organoids, which are still mini-organs, into systems that can be used for transplantation, as well as analyze design and disposition of cells and blood vessels in each organ to engineer 3D organ systems.

This team applies microdevice technologies for organoid studies. Currently, the organoid culture needs to have a perfusion system or to have a technology to control its morphology into 3D arbitrary shape. The team will try to develop a technology to achieve these functions using various organoids handled by other members and study how the organoid can be cultured or matured using the technology.

The team is targeting on developing 3D culture platform enabling initial cell cluster shape control and dynamic control of morphogen gradient surround cells. By employing cubic culture device as well as microfabrication technique, they will array cells into arbitrary 3D shape in a hydrogel. Subsequently, morphogen gradient will be controlled by integrating the cube device into the microfluidic chip, so that spatial-temporal information required to the body axis formation will be added to the cells. The team collaborates with Morimoto team to investigate the pattern formation of the tracheal mesenchyme organoid by employing the developed platform.

The team is aiming to apply our developed microfluidic devices for cellular micropatterning and long-term tissue culture techniques. We develop these techniques to studies promoted by core project members to create organoids. This will make it possible to analyze the structure and functions and long-term analysis. Thus, we aim to extend the feasibility of our techniques and also contribute to the development of organoid study.

Our team aims to design and construct an optimized extracellular matrix (ECM) environment for organoid formation. Using mammalian skin as a model, we will map the molecular composition of the ECM around each skin cell type. We will also develop an efficient purification system for different ECM molecules, and then combine these natural ECMs with artificial matrix substrates to construct and manipulate the ECM environment of organoids in a 4D manner.

The team is aiming to generate a whole lower urinary tract, including the kidney and bladder, using human iPS cells. To do so, they are working to elucidate the mechanism regulating mesodermal cell differentiation and the patterning of endoderm-derived intestinal tube formation, and develop approaches to control these mechanisms. They are examining tissue vascularization mechanisms and applying their knowledge to develop technology for maintaining culture of 3D organs. They also aim to use kidney organoids to mimic and measure physiological renal functions in vitro.

The team is aiming to generate a whole lower urinary tract, including the kidney and bladder, using human iPS cells. To do so, they are working to elucidate the mechanism regulating mesodermal cell differentiation and the patterning of endoderm-derived intestinal tube formation, and develop approaches to control these mechanisms. They are examining tissue vascularization mechanisms and applying their knowledge to develop technology for maintaining culture of 3D organs. They also aim to use kidney organoids to mimic and measure physiological renal functions in vitro.

The team is working to establish a new approach for self-organization-based induction of primordial organs and organoids, referred to as ‘direct reprogramming organogenesis.’ In addition, they are also striving to understand the mechanisms underlying organogenesis, particularly the molecular mechanisms inducing periodic morphological changes of hair follicles. They aim to cultivate primordial organs and organoids, which are still mini-organs, into systems that can be used for transplantation, as well as analyze design and disposition of cells and blood vessels in each organ to engineer 3D organ systems.

The team is aiming to develop an on-chip assay method to co-culture human iPSC-derived organoids and vasculature, based on a technology to create vascular network in a microfluidic device. It recapitulates an in vivo transplantation of organoids in vitro, which enables us to study tissue vascularization mechanisms on a chip. Furthermore, the team aims to mature organoids by long-term culture with perfusion through vasculature, and to apply the assay method to drug screening.

The team is working on generating alveolar organoids from isolated mouse alveolar stem cells, as well as tracheal mesenchymal organoids from ES and iPS cells. Using alveolar organoids, they are carrying out research on stemness and heterogeneity of tissue stem cells, as well as research of disease models. With tracheal mesenchyme organoids, they are developing 3D culture methods for inducing cartilage tissue structures and aiming to understand mechanisms of tracheal cartilage patterning.

The team is developing transplantation therapy using human iPS-derived retinal tissues (retinal pigment epithelium and photoreceptors) and gene therapy for the treatment of retinal degeneration diseases. To do so, they are improving the quality and culture methods of iPS cells and retinal organoids. Additionally, in order to constantly supply high-quality cells, which are currently produced by a handful of experienced researchers, a new technology is now being developed by combining the general-purpose humanoid robot, “Maholo”, and AI.

The project aims to develop standardized methods for analyzing the formation, degeneration and restoration of human brain tissues. Combining 3D brain organoid cultures of human pluripotent stem cells, 4D imaging and image analysis, we establish efficient methods to faithfully recapitulate and to quantitatively analyze the ontogenetic formation of human brain tissues. We induce degeneration of the formed tissues to construct human neurological disease models. We further explore the ways to prevent or restore the degeneration for clinical treatment and drug discovery.

Upon requests from the project members, the team collectively supports the Organoid Project in introducing genetic modifications to human iPS/ES cells. Genetically engineered human iPS/ES cells are extensively used for the generation of organoids in affiliated research projects. We will develop an improved platform to produce genetically engineered human iPS/ES cells by establishing culture protocols that can render robust undifferentiated state and high pluripotency and by optimizing gene modification methods.

The team is developing transplantation therapy using human iPS-derived retinal tissues (retinal pigment epithelium and photoreceptors) and gene therapy for the treatment of retinal degeneration diseases. To do so, they are improving the quality and culture methods of iPS cells and retinal organoids. Additionally, in order to constantly supply high-quality cells, which are currently produced by a handful of experienced researchers, a new technology is now being developed by combining the general-purpose humanoid robot, “Maholo”, and AI.

Using our developed automated systems such as a single-cell picking robot, we will analyze features of cells from a tissue, and will identify a cell which exhibits higher stemness potential. We will also perform whole genome expression analysis to understand states of these cells, and to find molecular markers. These approaches will contribute to efficient formation of organoids from tissue stem cells, and to in vivo studies as well.