Kyushu University Academic Staff Educational and Research Activities Database
List of Papers
Takashi Miura Last modified date:2022.01.19

Professor / Department of Anatomy and Cell Biology / Department of Basic Medicine / Faculty of Medical Sciences

1. Rei Mukomoto, Yuji Nashimoto, Takato Terai, Takuto Imaizumi, Kaoru Hiramoto, Kosuke Ino, Ryuji Yokokawa, Takashi Miura, Hitoshi Shiku, Oxygen consumption rate of tumour spheroids during necrotic-like core formation., The Analyst, 10.1039/d0an00979b, 2020.07, Hypoxia is one of the major hallmarks of solid tumours and is associated with the poor prognosis of various cancers. A multicellular aggregate, termed a spheroid, has been used as a tumour model with a necrotic-like core for more than 45 years. Oxygen metabolism in spheroids has been studied using phosphorescence quenching and oxygen-sensitive electrodes. However, these conventional methods require chemical labelling and physical insertion of the electrode into each spheroid, which may be functionally and structurally disruptive. Scanning electrochemical microscopy (SECM) can non-invasively analyse oxygen metabolism. Here, we used SECM to investigate whether the changes of the internal structure of spheroids affect the oxygen metabolism. We investigated the oxygen consumption rate (OCR) of MCF-7 breast tumour spheroids with and without a necrotic-like core. A numerical simulation was used to describe a method for estimating the OCR of spheroids that settled at the bottom of the conventional culture plates. The OCR per spheroid volume decreased with increasing spheroid radius, indicating the limitation of the oxygen supply to the core of the MCF-7 spheroid. Formation of the necrotic-like core did not affect the oxygen metabolism significantly, implying that the core had minimal contribution to the OCR even before necrosis occurred. OCR analysis using SECM non-invasively monitors the change of oxygen metabolism in tumour spheroids. The approach is promising to evaluate various three-dimensional culture models..
2. Satoru Okuda, Takashi Miura, Yasuhiro Inoue, Taiji Adachi, Mototsugu Eiraku, Combining Turing and 3D vertex models reproduces autonomous multicellular morphogenesis with undulation, tubulation, and branching, SCIENTIFIC REPORTS, 10.1038/s41598-018-24858-2, 8, 1, 2386, 2018.06.
3. Tomoyasu Shinoda, Arata Nagasaka, Yasuhiro Inoue, Ryo Higuchi, Yoshiaki Minami, Kagayaki Kato, Makoto Suzuki, Takefumi Kondo, Takumi Kawaue, Kanako Saito, Naoto Ueno, Yugo Fukazawa, Masaharu Nagayama, Takashi Miura, Taiji Adachi, Takaki Miyata, Elasticity-based boosting of neuroepithelial nucleokinesis via indirect energy transfer from mother to daughter, PLOS BIOLOGY, 10.1371/journal.pbio.2004426, 16, 4, 2018.04, Neural progenitor cells (NPCs), which are apicobasally elongated and densely packed in the developing brain, systematically move their nuclei/somata in a cell cycle-dependent manner, called interkinetic nuclear migration (IKNM): apical during G2 and basal during G1. Although intracellular molecular mechanisms of individual IKNM have been explored, how heterogeneous IKNMs are collectively coordinated is unknown. Our quantitative cell-biological and in silico analyses revealed that tissue elasticity mechanically assists an initial step of basalward IKNM. When the soma of an M-phase progenitor cell rounds up using actomyosin within the subapical space, a microzone within 10 mu m from the surface, which is compressed and elastic because of the apical surface's contractility, laterally pushes the densely neighboring processes of non-M-phase cells. The pressed processes then recoil centripetally and basally to propel the nuclei/somata of the progenitor's daughter cells. Thus, indirect neighbor-assisted transfer of mechanical energy from mother to daughter helps efficient brain development..
4. Engineering a Perfusable Vascular Network in a Microfluidic Device for a Morphological Analysis.
5. Yuji Nashimoto, Tomoya Hayashi, Itsuki Kunita, Akiko Nakamasu, Yu-Suke Torisawa, Masamune Nakayama, Hisako Takigawa-Imamura, Hidetoshi Kotera, Koichi Nishiyama, Takashi Miura, Ryuji Yokokawa, Integrating perfusable vascular networks with a three-dimensional tissue in a microfluidic device., Integrative biology : quantitative biosciences from nano to macro, 10.1039/c7ib00024c, 9, 6, 506-518, 2017.06, Creating vascular networks in tissues is crucial for tissue engineering. Although recent studies have demonstrated the formation of vessel-like structures in a tissue model, long-term culture is still challenging due to the lack of active perfusion in vascular networks. Here, we present a method to create a three-dimensional cellular spheroid with a perfusable vascular network in a microfluidic device. By the definition of the cellular interaction between human lung fibroblasts (hLFs) in a spheroid and human umbilical vein endothelial cells (HUVECs) in microchannels, angiogenic sprouts were induced from microchannels toward the spheroid; the sprouts reached the vessel-like structures in a spheroid to form a continuous lumen. We demonstrated that the vascular network could administer biological substances to the interior of the spheroid. As cell density in the spheroid is similar to that of a tissue, the perfusable vasculature model opens up new possibilities for a long-term tissue culture in vitro..
6. Yuji Nashimoto, Itsuki Kunita, Akiko Nakamasu, Yu-suke Torisawa, Masamune Nakayama, Hidetoshi Kotera, Koichi Nishiyama, Takashi Miura, Ryuji Yokokawa, ENGINEERING A THREE-DIMENSIONAL TISSUE MODEL WITH A PERFUSABLE VASCULATURE IN A MICROFLUIDIC DEVICE, 30TH IEEE INTERNATIONAL CONFERENCE ON MICRO ELECTRO MECHANICAL SYSTEMS (MEMS 2017), 10.1109/MEMSYS.2017.7863476, 592-595, 2017.04, In this study, we developed a microfluidic platform for a three-dimensional tissue model with a perfusable capillary network, which will allow, for the first time, a perfusion-culture in a tissue model with a high cell density. Our group previously reported that a spheroid of lung fibroblasts induced angiogenic sprouts from microchannels [1]. In this study, we successfully connected angiogenic sprouts to the vessel-like hollow structure in a spheroid and perfused the formed vascular network through microfluidic channels to the spheroid. This model opens up new techniques for tissue-culture for long-term..
7. Theoretical Models of Vascular Pattern Formation..
8. Takumi Higaki, Natsumaro Kutsuna, Kae Akita, Hisako Takigawa-Imamura, Kenji Yoshimura, Takashi Miura, A Theoretical Model of Jigsaw-Puzzle Pattern Formation by Plant Leaf Epidermal Cells, PLOS COMPUTATIONAL BIOLOGY, 10.1371/journal.pcbi.1004833, 12, 4, e1004833, 2016.04, Plant leaf epidermal cells exhibit a jigsaw puzzle-like pattern that is generated by interdigitation of the cell wall during leaf development. The contribution of two ROP GTPases, ROP2 and ROP6, to the cytoskeletal dynamics that regulate epidermal cell wall interdigitation has already been examined; however, how interactions between these molecules result in pattern formation remains to be elucidated. Here, we propose a simple interface equation model that incorporates both the cell wall remodeling activity of ROP GTPases and the diffusible signaling molecules by which they are regulated. This model successfully reproduces pattern formation observed in vivo, and explains the counterintuitive experimental results of decreased cellulose production and increased thickness. Our model also reproduces the dynamics of three-way cell wall junctions. Therefore, this model provides a possible mechanism for cell wall interdigitation formation in vivo..
9. Tomoya Hayashi, Hisako Takigawa-Imamura, Koichi Nishiyama, Hirofumi Shintaku, Hidetoshi Kotera, Takashi Miura, Ryuji Yokokawa, VASCULAR NETWORK FORMATION FOR A LONG-TERM SPHEROID CULTURE BY CO-CULTURING ENDOTHELIAL CELLS AND FIBROBLASTS, 2015 28TH IEEE INTERNATIONAL CONFERENCE ON MICRO ELECTRO MECHANICAL SYSTEMS (MEMS 2015), 476-479, 2015.04, In this paper, we present a poly-dimethylsiloxane (PDMS) microfluidic device to create a vascular network for a long-term spheroid culture, of which network and spheroid are consist of human umbilical vein endothelial cells (HUVEC) and normal human lung fibroblasts (LF), respectively. Following device design, fabrication, and fundamental evaluation of HUVEC sprouting conditions, we visualized that HUVEC networks were successfully formed by the co-culture with LFs and reached a LF-based spheroid. Moreover, perfusability of the network was evaluated by injecting fluorescent microbeads. This platform will be applicable for long-term tissue cultures to understand morphogenesis and modeling of blood vessel functions..
10. Takashi Miura, Models of lung branching morphogenesis, Journal of Biochemistry, 10.1093/jb/mvu087, 157, 3, 121-127, 2015.03, Vertebrate airway has a tree-like-branched structure. This structure is generated by repeated tip splitting, which is called branching morphogenesis. Although this phenomenon is extensively studied in developmental biology, the mechanism of the pattern formation is not well understood. Conversely, there are many tree-like structures in purely physical or chemical systems, and their pattern formation mechanisms are well-understood using mathematical models. Recent studies correlate these biological observations and mathematical models to understand lung branching morphogenesis. These models use slightly different mechanisms. In this article, we will review recent progress in modelling lung branching morphogenesis, and future directions to experimentally verify the models..
11. Ryuzo Arai, Masahiko Kobayashi, Hideto Harada, Hiroyuki Tsukiyama, Takahiko Saji, Yoshinobu Toda, Yoshihiro Hagiwara, Takashi Miura, Shuichi Matsuda, Anatomical study for SLAP lesion repair, Knee Surgery, Sports Traumatology, Arthroscopy, 10.1007/s00167-013-2385-3, 22, 2, 435-441, 2014.02, Purpose: The purpose of this study was to meticulously observe the structures around the origin of the long head of the biceps tendon (LHB) in order to propose a method of anatomical superior labrum anterior and posterior repair. Methods: Twenty-eight shoulders of 16 cadavers with intact LHB origin were macroscopically investigated. Among them, 20 shoulders with an intact superior labrum were additionally observed, to determine whether the anterior edge of LHB on the labrum (point 'A') was anterior to the supraglenoid tubercle. Serial sections vertical to LHB were observed using ordinary light and polarized microscopy in three glenoids and scanning acoustic microscopy in one. Results: The labrum had a meniscal appearance, and no LHB fibre was sent anterior to the anterior edge of the supraglenoid tubercle. 'A' was not located more posterior than the supraglenoid tubercle. All specimens had the so-called 'the sheet-like structure', in which the portion closer to the LHB origin tends to be stiffer. Fibres of the sheet-like structure ran vertically to LHB. Conclusion: Fibre orientation and the stiffness of the sheet-like structure suggest its support of LHB. As LHB fibres do not anteriorly cross over 'A', 'A' could be a landmark for the anterior border of LHB, independent from the sheet-like structure. Considering a previous report mentioning that the horizontal mattress suture maintains the meniscus-like structure which might be sufficient for proper motion of the normal superior labrum, the horizontal mattress suture not crossing over 'A' should be recommended from the viewpoint of functional anatomy. © 2013 Springer-Verlag Berlin Heidelberg..
12. Miura, T., Hartmann, D., Kinboshi, M., Komada, M., Ishibashi, M., Shiota, K., The cyst-branch difference in developing chick lung results from a different morphogen diffusion coefficient, Mechanisms of Development, 10.1016/j.mod.2008.11.006, 126, 3-4, 160-172, 2009.04, The developing avian lung is formed mainly by branching morphogenesis, but there is also a unique cystic structure, the air sac, in the ventral region. It has been shown that mesenchymal tissue is responsible for the differential development of a cystic or branched structure, and that the transcription factor Hoxb may be involved in determining this regional difference. We have previously developed two scenarios for branch-cyst transition, both experimentally and theoretically: increased production or increased diffusion of FGF. The aim of the present study was to discover whether one of these scenarios actually operates in the ventral region of the chick lung. We found that the FGF10 level was lower while the diffusion of FGF10 was more rapid in the ventral lung, indicating that the second scenario is more plausible. There are two possibilities as to why the diffusion of FGF10 differs between the two regions: (1) diffusion is facilitated by the looser tissue organisation of the ventral lung mesenchyme; (2) stronger expression of heparan sulphate proteoglycan ( HSPG) in the dorsal lung traps FGF and decreases the effective diffusion coefficient. Mathematical analysis showed that the dorsal-ventral difference in the amount of HSPG is not sufficient to generate the observed difference in pattern, indicating that both extracellular matrix and tissue architecture play a role in this system. These results suggest that the regional cystic-branched difference within the developing chick lung results from a difference in the rate of diffusion of morphogen between the ventral and dorsal regions due to differential levels of HSPG and a different mesenchymal structure. (C) 2008 Elsevier Ireland Ltd. All rights reserved..
13. T. Miura, R. Tanaka, In vitro vasculogenesis models revisited - Measurement of VEGF diffusion in matrigel, Mathematical Modelling of Natural Phenomena, 10.1051/mmnp/20094404, 4, 4, 118-130, 2009.01, The circulatory system is one of the first to function during development. The earliest event in the system's development is vasculogenesis, whereby vascular progeniter cells form clusters called blood islands, which later fuse to form capillary networks. There exists a very good in vitro system that mimics this process. When HUVECs (Human Umbilical Vein Endothelial Cells) are cultured on Matrigel, they spontaneously form a capillary network structure. Two theoretical models have been proposed to explain the pattern formation of this in vitro system. Both models utilize chemotaxis to generate spatial instability, and one model specifies VEGF as the chemoattractant. However, there are several unknown factors concerning the experimental model. First, the pattern formation process occurs at the interface between the liquid medium and Matrigel, and it is unclear whether diffusion in the liquid or gel is critical. Second, the diffusion coefficient of VEGF, which determines the spatial scale of the capillary structure, has not been properly measured. In the present study, we modified the experimental system to clarify the effect of diffusion in Matrigel, and experimentally measured the diffusion coefficient of VEGF in this system. The relationship with the spatial scale of the pattern generated is discussed. © EDP Sciences, 2009..
14. [Turing reaction-diffusion model in developmental biology: application to limb skeletal pattern formation]..
15. Atsushi Hinoue, Toshiya Takigawa, Takashi Miura, Yoshihiko Nishimura, Shigehiko Suzuki, Kohei Shiota, Disruption of actin cytoskeleton and anchorage-dependent cell spreading induces apoptotic death of mouse neural crest cells cultured in vitro, Anatomical Record - Part A Discoveries in Molecular, Cellular, and Evolutionary Biology, 10.1002/ar.a.20150, 282, 2, 130-137, 2005.02, In vertebrate embryos, neural crest cells emigrate out of the neural tube and contribute to the formation of a variety of neural and nonneural tissues. Some neural crest cells undergo apoptotic death during migration, but its biological significance and the underlying mechanism are not well understood. We carried out an in vitro study to examine how the morphology and survival of cranial neural crest (CNC) cells of the mouse embryo are affected when their actin cytoskeleton or anchorage-dependent cell spreading is perturbed. Disruption of actin fiber organization by cytochalasin D (1 μg/ml) and inhibition of cell attachment by matrix metalloproteinase-2 (MMP-2
2.0 units/ml) were followed by morphologic changes and apoptotic death of cultured CNC cells. When the actin cytoskeleton was disrupted by cytochalasin D, the morphologic changes of cultured CNC cells preceded DNA fragmentation. These results indicate that the maintenance of cytoskeleton and anchorage-dependent cell spreading are required for survival of CNC cells. The spatially and temporally regulated expression of proteinases may be essential for the differentiation and migration of neural crest cells. © 2005 Wiley-Liss, Inc..
16. Takashi Miura, Philip K. Maini, Periodic pattern formation in reaction-diffusion systems: An introduction for numerical simulation, Anatomical Science International, 10.1111/j.1447-073x.2004.00079.x, 79, 3, 112-123, 2004.09, The aim of the present review is to provide a comprehensive explanation of Turing reaction-diffusion systems in sufficient detail to allow readers to perform numerical calculations themselves. The reaction-diffusion model is widely studied in the field of mathematical biology, serves as a powerful paradigm model for self-organization and is beginning to be applied to actual experimental systems in developmental biology. Despite the increase in current interest, the model is not well understood among experimental biologists, partly because appropriate introductory texts are lacking. In the present review, we provide a detailed description of the definition of the Turing reaction-diffusion model that is comprehensible without a special mathematical background, then illustrate a method for reproducing numerical calculations with Microsoft Excel. We then show some examples of the patterns generated by the model. Finally, we discuss future prospects for the interdisciplinary field of research involving mathematical approaches in developmental biology..
17. Takashi Miura, Kohei Shiota, TGFβ2 acts as an 'activator' molecule in reaction-diffusion model and is involved in cell sorting phenomenon in mouse limb micromass culture, Developmental Dynamics, 10.1002/(SICI)1097-0177(200003)217:3<241::AID-DVDY2>3.0.CO;2-K, 217, 3, 241-249, 2000.04, It was previously speculated that TGFβ acts as an 'activator'-molecule in chondrogenic pattern formation in the limb micromass culture system, but its precise role and relationship with the cell sorting phenomenon have not been properly studied. In the present study, we examined whether the TGFβ2 molecule satisfies the necessary conditions for an 'activator'-molecule in the reaction-diffusion model. Firstly, we showed that TGFβ2 became localized at chondrogenic sites during the establishment of a chondrogenic pattern, and exogenous TGFβ2 promoted chondrogenesis when added in the culture medium. Secondly, TGFβ2 protein was shown to promote the production of its own mRNA after 3 hr, indicating that a positive feedback mechanism exists which may be responsible for the emergence of the chondrogenic pattern. We then found that when locally applied with beads, TGFβ2 suppressed chondrogenesis around the beads, indicating it induces the lateral inhibitory mechanism, which is a key element for the formation of the periodic pattern. We also examined the possible effects of TGFβ2 on the cell sorting phenomenon and found that TGFβ2 exerts differential chemotactic activity on proximal and distal mesenchyme cells of the limb bud, and at very early phases of differentiation TGFβ2 promotes the expression of N-cadherin protein which is known to be involved in pattern formation in this culture system. These findings suggest that TGFβ2 acts as an 'activator'-like molecule in chondrogenic pattern formation in vitro, and is possibly responsible for the cell sorting phenomenon. (C) 2000 Wiley- Liss, Inc..
18. Takashi Miura, Kohei Shiota, Time-lapse observation of branching morphogenesis of the lung bud epithelium in mesenchyme-free culture and its relationship with the localization of actin filaments, International Journal of Developmental Biology, 44, 8, 899-902, 2000.04, It has been shown that branching morphogenesis of the lung bud is mediated by epithelial-mesenchymal interaction via such molecules as FGF10, BMP4 and Shh. However, a recent study showed that the isolated lung epithelium still undergoes branching morphogenesis in vitro even in the absence of mesenchyme (Nogawa and Ito, 1995). In the present study, we observed in vitro the dynamic movement of the isolated lung epithelium of the fetal mouse using time-lapse recording, and investigated the roles of actin filaments in branching of the lung bud. First, time-lapse observation of the initial phase of lung branching morphogenesis revealed that at the sites of cleft formation, the epithelial surface was retracted inward from its original position. From this observation we assumed that there should be some structures which exert a physical force on the epithelium, and the localization and arrangement of actin fibers in the cultured lung epithelium were examined at various stages of branching morphogenesis. At the prebudding (6 h) and onset-budding (24 h) stages, no specific localization of actin filaments was observed in the lung bud epithelium, but at the postbudding stage (48 h) they were localized densely in the cells at the tip of the branched lung epithelium. The cell density was not different between the tip and cleft regions of the lung bud epithelium. When cultured with FGF-soaked beads, an actin-rich region was induced at the tip of the lung bud which was growing toward an FGF-soaked bead. These results indicate that actin fibers do not play a significant part in cleft formation but can be secondarily induced by FGF in the surrounding matrix and play some roles at later shaping of the branch in lung morphogenesis..