記述言語：英語   掲載種別：研究論文（学術雑誌）  

<title>Abstract</title>Microglia are resident immune cells in the central nervous system, showing a regular distribution. Advancing microscopy and image processing techniques have contributed to elucidating microglia’s morphology, dynamics, and distribution. However, the mechanism underlying the regular distribution of microglia remains to be elucidated. First, we quantitatively confirmed the regularity of the distribution pattern of microglial soma in the retina. Second, we formulated a mathematical model that includes factors that may influence regular distribution. Next, we experimentally quantified the model parameters (cell movement, process formation, and ATP dynamics). The resulting model simulation from the measured parameters showed that direct cell–cell contact is most important in generating regular cell spacing. Finally, we tried to specify the molecular pathway responsible for the repulsion between neighboring microglia.

DOI： 10.1038/s41598-021-01820-3

記述言語：英語   掲載種別：研究論文（学術雑誌）  

In vertebrates, sperm is generated in testicular tube-like structures called seminiferous tubules. The differentiation stages of spermatogenesis exhibit a dynamic spatiotemporal wavetrain pattern. There are two types of pattern-the vertical type, which is observed in mice, and the helical type, which is observed in humans. The mechanisms of this pattern difference remain little understood. In the present study, we used a three-species reaction-diffusion model to reproduce the wavetrain pattern observed in vivo. We hypothesized that the wavelength of the pattern in mice was larger than that in humans and undertook numerical simulations. We found complex patterns of helical and vertical pattern frequency, which can be understood by pattern selection using boundary conditions. From these theoretical results, we predicted that a small number of vertical patterns should be present in human seminiferous tubules. We then found vertical patterns in histological sections of human tubules, consistent with the theoretical prediction. Finally, we showed that the previously reported irregularity of the human pattern could be reproduced using two factors: a wider unstable wavenumber range and the irregular geometry of human compared with mouse seminiferous tubules. These results show that mathematical modeling is useful for understanding the pattern dynamics of seminiferous tubules in vivo.

DOI： 10.1007/s11538-021-00863-x

記述言語：英語   掲載種別：研究論文（学術雑誌）  

A lack of perfusion has been one of the most significant obstacles for three-dimensional culture systems of organoids and embryonic tissues. Here, we developed a simple and reliable method to implement a perfusable capillary network in vitro. The method employed the self-organization of endothelial cells to generate a capillary network and a static pressure difference for culture medium circulation, which can be easily introduced to standard biological laboratories and enables long-term cultivation of vascular structures. Using this culture system, we perfused the lumen of the self-organized capillary network and observed a flow-induced vascular remodeling process, cell shape changes, and collective cell migration. We also observed an increase in cell proliferation around the self-organized vasculature induced by flow, indicating functional perfusion of the culture medium. We also reconstructed extravasation of tumor and inflammatory cells, and circulation inside spheroids including endothelial cells and human lung fibroblasts. In conclusion, this system is a promising tool to elucidate the mechanisms of various biological processes related to vascular flow.

DOI： 10.1371/journal.pone.0240552

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Pericytes (PCs) wrap around endothelial cells (ECs) and perform diverse functions in physiological and pathological processes. Although molecular interactions between ECs and PCs have been extensively studied, the morphological processes at the cellular level and their underlying mechanisms have remained elusive. In this study, using a simple cellular Potts model, we explored the mechanisms for EC wrapping by PCs. Based on the observed
<italic>in vitro</italic>
cell wrapping in three-dimensional PC–EC coculture, the model identified four putative contributing factors: preferential adhesion of PCs to the extracellular matrix (ECM), strong cell–cell adhesion, PC surface softness and larger PC size. While cell–cell adhesion can contribute to the prevention of cell segregation and the degree of cell wrapping, it cannot determine the orientation of cell wrapping alone. While atomic force microscopy revealed that PCs have a larger Young’s modulus than ECs, the experimental analyses supported preferential ECM adhesion and size asymmetry. We also formulated the corresponding energy minimization problem and numerically solved this problem for specific cases. These results give biological insights into the role of PC–ECM adhesion in PC coverage. The modelling framework presented here should also be applicable to other cell wrapping phenomena observed
<italic>in vivo</italic>
.

DOI： 10.1098/rsif.2019.0739

記述言語：英語   掲載種別：研究論文（学術雑誌）  

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.

DOI： 10.1039/c7ib00024c

記述言語：その他   掲載種別：研究論文（学術雑誌）  

DOI： 10.1093/pcp/pcw183

記述言語：英語   掲載種別：研究論文（学術雑誌）  

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.

DOI： 10.1371/journal.pcbi.1004833

記述言語：その他   掲載種別：研究論文（学術雑誌）  

DOI： 10.1088/1478-3975/10/6/066007

記述言語：その他   掲載種別：研究論文（学術雑誌）  

DOI： 10.1126/science.1179047

記述言語：その他   掲載種別：研究論文（学術雑誌）  

DOI： 10.1111/j.1469-7580.2009.01148.x

記述言語：英語   掲載種別：研究論文（学術雑誌）  

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.

DOI： 10.1016/j.mod.2008.11.006

記述言語：その他   掲載種別：研究論文（学術雑誌）  

DOI： 10.1016/j.jtbi.2006.05.009

記述言語：その他   掲載種別：研究論文（学術雑誌）  

DOI： 10.1016/j.jtbi.2005.10.016

記述言語：その他   掲載種別：研究論文（学術雑誌）  

DOI： 10.1016/j.bulm.2003.09.009

記述言語：その他   掲載種別：研究論文（学術雑誌）  

DOI： 10.1016/s0925-4773(02)00132-6

記述言語：英語   掲載種別：研究論文（学術雑誌）  

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.

DOI： 10.1002/(SICI)1097-0177(200003)217:3<241::AID-DVDY2>3.0.CO;2-K

記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Cancer Science  

Metastasis to the liver is a leading cause of death in patients with colorectal cancer. To investigate the characteristics of cancer cells prone to metastasis, we utilized an isogenic model of BALB/c and colon tumor 26 (C26) cells carrying an active KRAS mutation. Liver metastatic (LM) 1 cells were isolated from mice following intrasplenic transplantation of C26 cells. Subsequent injections of LM1 cells generated LM2 cells, and after four cycles, LM4 cells were obtained. In vitro, using a perfusable capillary network system, we found comparable extravasation frequencies between C26 and LM4 cells. Both cell lines showed similar growth rates in vitro. However, C26 cells showed higher glucose consumption, whereas LM4 cells incorporated more fluorescent fatty acids (FAs). Biochemical analysis revealed that LM4 cells had higher cholesterol levels than C26 cells. A correlation was observed between fluorescent FAs and cholesterol levels detected using filipin III. LM4 cells utilized FAs as a source for cholesterol synthesis through acetyl-CoA metabolism. In cellular analysis, cholesterol accumulated in punctate regions, and upregulation of NLRP3 and STING proteins, but not mTOR, was observed in LM4 cells. Treatment with a cholesterol synthesis inhibitor (statin) induced LM4 cell death in vitro and suppressed LM4 cell growth in the livers of nude mice. These findings indicate that colorectal cancer cells prone to liver metastasis show cholesterol-dependent growth and that statin therapy could help treat liver metastasis in immunocompromised patients.

DOI： 10.1111/cas.16331

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記述言語：その他   掲載種別：研究論文（学術雑誌）  

Abstract

Tooth development is governed largely by epithelial–mesenchymal interactions and is mediated by numerous signaling pathways. This type of morphogenetic processes has been explained by reaction–diffusion systems, especially in the framework of a Turing model. Here we focus on morphological and developmental differences between upper and lower molars in mice by modeling 2D pattern formation in a Turing system. Stripe vs. spot patterns are the primary types of variation in a Turing model. We show that the complexity of the cusp cross-sections can distinguish between stripe vs. spot patterns, and mice have stripe-like upper and spot-like lower molar morphologies. Additionally, our computational modeling that incorporates empirical data on tooth germ growth traces the order of cusp formation and relative position of the cusps in upper and lower molars in mice. We further propose a hypothetical framework of developmental mechanism that could help us understand the evolution of the highly variable nature of mammalian molars associated with the acquisition of the hypocone and the increase of lophedness.

DOI： 10.1038/s41598-022-13539-w

記述言語：その他   掲載種別：研究論文（学術雑誌）  

DOI： 10.1016/j.bios.2022.114808

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Angiogenesis is regulated in coordinated fashion by chemical and mechanical cues acting on endothelial cells (ECs). However, the mechanobiological mechanisms of angiogenesis remain unknown. Herein, we demonstrate a crucial role of blood flow-driven intraluminal pressure (IP) in regulating wound angiogenesis. During wound angiogenesis, blood flow-driven IP loading inhibits elongation of injured blood vessels located at sites upstream from blood flow, while downstream injured vessels actively elongate. In downstream injured vessels, F-BAR proteins, TOCA1 and CIP4, localize at leading edge of ECs to promote N-WASP-dependent Arp2/3 complex-mediated actin polymerization and front-rear polarization for vessel elongation. In contrast, IP loading expands upstream injured vessels and stretches ECs, preventing leading edge localization of TOCA1 and CIP4 to inhibit directed EC migration and vessel elongation. These data indicate that the TOCA family of F-BAR proteins are key actin regulatory proteins required for directed EC migration and sense mechanical cell stretching to regulate wound angiogenesis.

DOI： 10.1038/s41467-022-30197-8

記述言語：その他   掲載種別：研究論文（学術雑誌）  

<title>Abstract</title>The median palatal suture serves as a growth center for the maxilla; inadequate growth at this site causes malocclusion and dental crowding. However, the pattern formation mechanism of palatal sutures is poorly understood compared with that of calvarial sutures such as the sagittal suture. In the present study, therefore, we compared the morphological characteristics of sagittal and palatal sutures in human bone specimens. We found that palatal suture width was narrower than sagittal suture width, and the interdigitation amplitude of the palatal suture was lower than that of the sagittal suture. These tendencies were also observed in the neonatal stage. However, such differences were not observed in other animals such as chimpanzees and mice. We also used a mathematical model to reproduce the differences between palatal and sagittal sutures. After an extensive parameter search, we found two conditions that could generate the difference in interdigitation amplitude and suture width: bone differentiation threshold <inline-formula><alternatives><tex-math>$$v_c$$</tex-math><mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML">
<mml:msub>
<mml:mi>v</mml:mi>
<mml:mi>c</mml:mi>
</mml:msub>
</mml:math></alternatives></inline-formula> and growth speed <italic>c</italic>. We discuss possible biological interpretations of the observed pattern difference and its cause.

DOI： 10.1038/s41598-021-88255-y

記述言語：その他   掲載種別：研究論文（学術雑誌）  

DOI： 10.1016/j.jtbi.2020.110496

記述言語：英語   掲載種別：研究論文（学術雑誌）  

<title>Abstract</title>High vascularization is a biological characteristic of glioblastoma (GBM); however, an <italic>in-vitro</italic> experimental model to verify the mechanism and physiological role of vasculogenesis in GBM is not well-established. Recently, we established a self-organizing vasculogenic model using human umbilical vein endothelial cells (HUVECs) co-cultivated with human lung fibroblasts (hLFs). Here, we exploited this system to establish a realistic model of vasculogenesis in GBM. We developed two polydimethylsiloxane (PDMS) devices, a doughnut-hole dish and a 5-lane microfluidic device to observe the contact-independent effects of glioblastoma cells on HUVECs. We tested five patient-derived and five widely used GBM cell lines. Confocal fluorescence microscopy was used to observe the morphological changes in Red Fluorescent Protein (RFP)-HUVECs and fluorescein isothiocyanate (FITC)-dextran perfusion. The genetic and expression properties of GBM cell lines were analyzed. The doughnut-hole dish assay revealed KNS1451 as the only cells to induce HUVEC transformation to vessel-like structures, similar to hLFs. The 5-lane device assay demonstrated that KNS1451 promoted the formation of a vascular network that was fully perfused, revealing the functioning luminal construction. Microarray analysis revealed that KNS1451 is a mesenchymal subtype of GBM. Using a patient-derived mesenchymal GBM cell line, mature <italic>de-novo</italic> vessel formation could be induced in HUVECs by contact-independent co-culture with GBM in a microfluidic device. These results support the development of a novel in vitro research model and provide novel insights in the neovasculogenic mechanism of GBM and may potentially facilitate the future detection of unknown molecular targets.

DOI： 10.1007/s11033-020-06061-7

記述言語：英語   掲載種別：研究論文（学術雑誌）  

We found evidence of dynamic scaling in the spreading of Madin-Darby canine kidney (MDCK) cell monolayer, which can be characterized by the Hurst exponent α=0.86 and the growth exponent β=0.73, and theoretically and experimentally clarified the mechanism that governs the contour shape dynamics. Dynamic scaling refers to the roughness of the surface scales, both spatially and temporally. During the spreading of the monolayer, it is known that so-called leader cells generate the driving force and lead the other cells. Our time-lapse observations of cell behavior showed that these leader cells appeared at the early stage of the spreading and formed the monolayer protrusion. Informed by these observations, we developed a simple mathematical model that included differences in cell motility, cell-cell adhesion, and random cell movement. The model reproduced the quantitative characteristics obtained from the experiment, such as the spreading speed, the distribution of the increment, and the dynamic scaling law. Analysis of the model equation shows that the model can reproduce different scaling laws from (α=0.5,β=0.25) to (α=0.9,β=0.75), where the exponents α and β are determined by two dimensionless quantities determined by the microscopic cell behavior. From the analytical result, parameter estimation from the experimental results was achieved. The monolayer on the collagen-coated dishes showed a different scaling law, α=0.74,β=0.68, suggesting that cell motility increased ninefold. This result was consistent with the assay of the single-cell motility. Our study demonstrated that the dynamics of the contour of the monolayer were explained by the simple model, and we propose a mechanism that exhibits the dynamic scaling property.

DOI： 10.1103/physreve.102.062408

記述言語：その他   掲載種別：研究論文（学術雑誌）  

Sutures, the thin, soft tissue between skull bones, serve as the major craniofacial growth centers during postnatal development. In a newborn skull, the sutures are straight; however, as the skull develops, the sutures wind dynamically to form an interdigitation pattern. Moreover, the final winding pattern had been shown to have fractal characteristics. Although various molecules involved in suture development have been identified, the mechanism underlying the pattern formation remains unknown. In a previous study, we reproduced the formation of the interdigitation pattern in a mathematical model combining an interface equation and a convolution kernel. However, the generated pattern had a specific characteristic length, and the model was unable to produce a fractal structure with the model. In the present study, we focused on the anterior part of the sagittal suture and formulated a new mathematical model with time–space-dependent noise that was able to generate the fractal structure. We reduced our previous model to represent the linear dynamics of the centerline of the suture tissue and included a time–space-dependent noise term. We showed theoretically that the final pattern from the model follows a scaling law due to the scaling of the dispersion relation in the full model, which we confirmed numerically. Furthermore, we observed experimentally that stochastic fluctuation of the osteogenic signal exists in the developing skull, and found that actual suture patterns followed a scaling law similar to that of the theoretical prediction.

DOI： 10.1371/journal.pone.0235802

記述言語：英語   掲載種別：研究論文（学術雑誌）  

The development of retinal blood vessels has extensively been used as a model to study vascular pattern formation. To date, various quantitative measurements, such as size distribution have been performed, but the relationship between pattern formation mechanisms and these measurements remains unclear. In the present study, we first focus on the islands (small regions subdivided by the capillary network). We quantitatively measured the island size distribution in the retinal vascular network and found that it tended to exhibit an exponential distribution. We were able to recapitulate this distribution pattern in a theoretical model by implementing the stochastic disappearance of vessel segments around arteries could reproduce the observed exponential distribution of islands. Second, we observed that the diameter distribution of the retinal artery segment obeyed a power law. We theoretically showed that an equal bifurcation branch pattern and Murray's law could reproduce this pattern. This study demonstrates the utility of examining size distribution for understanding the mechanisms of vascular pattern formation.

DOI： 10.1371/journal.pone.0235373

記述言語：英語   掲載種別：研究論文（学術雑誌）  

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.

DOI： 10.1039/d0an00979b

記述言語：英語   掲載種別：研究論文（学術雑誌）  

<title>Abstract</title>A lack of microcirculation has been one of the most significant obstacles for three-dimensional culture systems of organoids and embryonic tissues. Here, we developed a simple and reliable method to implement a perfusable capillary network in vitro. The method employed the self-organization of endothelial cells to generate a capillary network and a static pressure difference for culture medium circulation, which can be easily introduced to standard biological laboratories and enables long-term cultivation of vascular structures. Using this culture system, we perfused the lumen of the self-organized capillary network and observed a flow-induced vascular remodeling process, cell shape changes, and collective cell migration. We also observed an increase in cell proliferation around the synthetic vasculature induced by flow, indicating functional perfusion of the culture medium. We also reconstructed extravasation of tumor and inflammatory cells, and circulation inside spheroids including endothelial cells and human lung fibroblasts. In conclusion, this system is a promising tool to elucidate the mechanisms of various biological processes related to vascular flow.

DOI： 10.1101/2020.05.12.067165

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Tumor vasculature creates a hostile tumor microenvironment (TME) in vivo and nourishes cancers, resulting in cancer progression and drug resistance. To mimic the biochemical and biomechanical environments of tumors in vitro, several models integrated with a vascular network have been reported. However, the tumor responses to biochemical and biomechanical stimuli were evaluated under static conditions and failed to incorporate the effects of blood flow to tumors. In this study, we present a tumor-on-a-chip platform that enables the evaluation of tumor activities with intraluminal flow in an engineered tumor vascular network. The fibroblasts in the tumor spheroid induced angiogenic sprouts, which constructed a perfusable vascular network in a tumor spheroid. The perfusability of the engineered vascular network was preserved during the culture. Moreover, perfusion for over 24 h significantly increased the proliferation activities of tumor cells and decreased cell death in the spheroid. Drug administration under perfusion condition did not show the dose-dependent effects of anticancer drugs on tumor activities in contrast to the results under static conditions. Our results demonstrate the importance of flow in a vascular network for the evaluation of tumor activities in a drug screening platform.

DOI： 10.1016/j.biomaterials.2019.119547

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Tight junctions (TJs) establish the epithelial barrier and are thought to form a membrane fence to regulate epithelial polarity, although the roles of TJs in epithelial polarity remain controversial. Claudins constitute TJ strands in conjunction with the cytoplasmic scaffolds ZO-1 and ZO-2 and play pivotal roles in epithelial barrier formation. However, how claudins and other TJ membrane proteins cooperate to organize TJs remains unclear. Here, we systematically knocked out TJ components by genome editing and show that while ZO-1/ZO-2–deficient cells lacked TJ structures and epithelial barriers, claudin-deficient cells lacked TJ strands and an electrolyte permeability barrier but formed membrane appositions and a macromolecule permeability barrier. Moreover, epithelial polarity was disorganized in ZO-1/ZO-2–deficient cells, but not in claudin-deficient cells. Simultaneous deletion of claudins and a TJ membrane protein JAM-A resulted in a loss of membrane appositions and a macromolecule permeability barrier and in sporadic epithelial polarity defects. These results demonstrate that claudins and JAM-A coordinately regulate TJ formation and epithelial polarity.

DOI： 10.1083/jcb.201812157

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1038/s41598-018-24858-2

記述言語：その他   掲載種別：研究論文（学術雑誌）  

DOI： 10.1016/j.devcel.2018.04.010

記述言語：英語   掲載種別：研究論文（学術雑誌）  

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.

DOI： 10.1371/journal.pbio.2004426

記述言語：日本語   掲載種別：研究論文（学術雑誌）  

Engineering a Perfusable Vascular Network in a Microfluidic Device for a Morphological Analysis

DOI： 10.1541/ieejsmas.138.275

記述言語：英語   掲載種別：研究論文（学術雑誌）  

A spheroid (a multicellular aggregate) is regarded as a good model of living tissues in the human body. Despite the significant advancement in the spheroid cultures, a perfusable vascular network in the spheroids remains a critical challenge for long-term culture required to maintain and develop their functions, such as protein expressions and morphogenesis. The protocol presents a novel method to integrate a perfusable vascular network within the spheroid in a microfluidic device. To induce a perfusable vascular network in the spheroid, angiogenic sprouts connected to microchannels were guided to the spheroid by utilizing angiogenic factors from human lung fibroblasts cultured in the spheroid. The angiogenic sprouts reached the spheroid, merged with the endothelial cells co-cultured in the spheroid, and formed a continuous vascular network. The vascular network could perfuse the interior of the spheroid without any leakage. The constructed vascular network may be further used as a route for supply of nutrients and removal of waste products, mimicking blood circulation in vivo. The method provides a new platform in spheroid culture toward better recapitulation of living tissues.

DOI： 10.3791/57242

記述言語：その他   掲載種別：研究論文（学術雑誌）  

DOI： 10.1016/j.jtbi.2017.06.012

記述言語：英語   掲載種別：研究論文（その他学術会議資料等）  

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.

DOI： 10.1109/MEMSYS.2017.7863476

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Reelin is an essential glycoprotein for the establishment of the highly organized six-layered structure of neurons of the mammalian neocortex. Although the role of Reelin in the control of neuronal migration has been extensively studied at the molecular level, the mechanisms underlying Reelin-dependent neuronal layer organization are not yet fully understood. In this study, we directly showed that Reelin promotes adhesion among dissociated neocortical neurons in culture. The Reelin-mediated neuronal aggregation occurs in an N-cadherin–dependent manner, both in vivo and in vitro. Unexpectedly, however, in a rotation culture of dissociated neocortical cells that gradually reaggregated over time, we found that it was the neural progenitor cells [radial glial cells (RGCs)], rather than the neurons, that tended to form clusters in the presence of Reelin. Mathematical modeling suggested that this clustering of RGCs could be recapitulated if the Reelin-dependent promotion of neuronal adhesion were to occur only transiently. Thus, we directly measured the adhesive force between neurons and N-cadherin by atomic force microscopy, and found that Reelin indeed enhanced the adhesiveness of neurons to N-cadherin; this enhanced adhesiveness began to be observed at 30 min after Reelin stimulation, but declined by 3 h. These results suggest that Reelin transiently (and not persistently) promotes N-cadherin–mediated neuronal aggregation. When N-cadherin and stabilized β-catenin were overexpressed in the migrating neurons, the transfected neurons were abnormally distributed in the superficial region of the neocortex, suggesting that appropriate regulation of N-cadherin–mediated adhesion is important for correct positioning of the neurons during neocortical development.

DOI： 10.1073/pnas.1615215114

記述言語：その他   掲載種別：研究論文（学術雑誌）  

DOI： 10.1016/j.jtbi.2016.08.003

記述言語：その他  

Theoretical Models of Vascular Pattern Formation.

DOI： 10.15017/1787037

記述言語：その他   掲載種別：研究論文（学術雑誌）  

Notch-mediated lateral inhibition regulates binary cell fate choice, resulting in salt and pepper patterns during various developmental processes. However, how Notch signaling behaves in combination with other signaling systems remains elusive. The wave of differentiation in the <italic>Drosophila</italic> visual center or “proneural wave” accompanies Notch activity that is propagated without the formation of a salt and pepper pattern, implying that Notch does not form a feedback loop of lateral inhibition during this process. However, mathematical modeling and genetic analysis clearly showed that Notch-mediated lateral inhibition is implemented within the proneural wave. Because partial reduction in EGF signaling causes the formation of the salt and pepper pattern, it is most likely that EGF diffusion cancels salt and pepper pattern formation in silico and in vivo. Moreover, the combination of Notch-mediated lateral inhibition and EGF-mediated reaction diffusion enables a function of Notch signaling that regulates propagation of the wave of differentiation.

DOI： 10.1073/pnas.1602739113

記述言語：その他   掲載種別：研究論文（学術雑誌）  

DOI： 10.1111/dgd.12292

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Articular cartilage (AC) covers the surface of bones in joints and functions as a cushion against mechanical loading. The tissue contains abundant extracellular matrix (ECM), which mainly consists of proteoglycans (PG) and collagen (COL) fibres. The property of AC is gradually changing by ageing with gravity loading. To know the property change of AC by initial gravity loading during short period after birth, we performed histological assays and proteomics assay on the AC of the femoral condyle in knee joints of perinatal rats. The water content (&#37;) was significantly decreased in neonate AC compared with fetal AC. During the perinatal stages (E19 and P0), the localizations of glycosaminoglycan (GAG) and type I and II COLs were homogeneous. The density of chondrocytes was significantly decreased in the deeper layers comparing with the surface layer in neonate AC. In addition, we found a drastic change in the protein expression pattern on proteomic analysis. The expressions of ECM components were relatively increased in neonate AC compared with fetal AC.

DOI： 10.1111/ahe.12165

記述言語：英語   掲載種別：研究論文（その他学術会議資料等）  

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.

記述言語：英語   掲載種別：研究論文（学術雑誌）  

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.

DOI： 10.1093/jb/mvu087

記述言語：英語   掲載種別：研究論文（学術雑誌）  

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.

DOI： 10.1007/s00167-013-2385-3

記述言語：その他   掲載種別：研究論文（学術雑誌）  

DOI： 10.1126/scisignal.2004038

記述言語：その他   掲載種別：研究論文（学術雑誌）  

記述言語：その他   掲載種別：研究論文（学術雑誌）  

DOI： 10.1007/s00276-011-0840-8

記述言語：その他   掲載種別：研究論文（学術雑誌）  

記述言語：英語   掲載種別：研究論文（学術雑誌）  

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.

DOI： 10.1051/mmnp/20094404

記述言語：その他   掲載種別：研究論文（学術雑誌）  

DOI： 10.1242/dev.015891

記述言語：その他  

DOI： 10.1016/s0070-2153(07)81010-6

記述言語：その他   掲載種別：研究論文（学術雑誌）  

DOI： 10.1093/imammb/dql029

記述言語：その他  

[Turing reaction-diffusion model in developmental biology: application to limb skeletal pattern formation].

記述言語：英語   掲載種別：研究論文（学術雑誌）  

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.

DOI： 10.1002/ar.a.20150

記述言語：その他   掲載種別：研究論文（学術雑誌）  

DOI： 10.1111/j.0021-8782.2004.00335.x

記述言語：英語   掲載種別：研究論文（学術雑誌）  

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.

DOI： 10.1111/j.1447-073x.2004.00079.x

記述言語：英語  

Stage-dependency of the effect of ethanol on chondrogenic differentiation in mouse limb micromass culture

記述言語：その他   掲載種別：研究論文（学術雑誌）  

DOI： 10.1007/s004290050329

記述言語：英語   掲載種別：研究論文（学術雑誌）  

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.

記述言語：その他   掲載種別：研究論文（学術雑誌）  

DOI： 10.1002/(sici)1097-0185(20000101)258:1<100::aid-ar11>3.0.co;2-3

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Mechanism of scaling law formation in retina vasculature - modeling and experiment

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Pericyte coverage of endothelial cells: in vitro experiments and computational modeling.

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Modeling of skull suture pattern formation

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Mechanism of skull suture pattern formation

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Distribution pattern of immune cells - quantification and mathematical modeling of pattern formation mechanism

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Establishment of perfusable vascular network for understanding multicellular pattern formation

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Analyses of sizes of self-organized patterns composed by endothelial cell

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Modeling vascular scaling law formation in retina vasculature

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Skull suture and plant cell wall: Mechanism of interdigitation pattern formation

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Mechanism of scaling law formation in retina vasculature - modeling and experiment

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A Mathematical model to generate fractal structure in skull suture

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Mechanism of skull suture pattern formation

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Pericyte coverage of endothelial cells: in vitro experiments and computational modeling.

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Modeling of skull suture pattern formation

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Distribution pattern of immune cells - quantification and mathematical modeling of pattern formation mechanism

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Establishment of perfusable vascular network for understanding multicellular pattern formation

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Perfusable synthetic self-organized vascular network - experiment and modeling

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Mechanisms of Pattern Formation in Limb Bud Micromass Culture: Possible Relationship with in vivo Pattern Formation

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Morphogenesis of Embryonic Lung Epithelium in Vitro: Possible Relationship with Theoretical Models

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Role of actin filaments in branching morphogenesis of the embryonic lung epithelium in vitro

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Speed of pattern appearance in reaction-diffusion system

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Modelling dynamics of skull suture pattern formation during development

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Cyst-branch difference in chick lung

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Mechanism of lung branching morphogenesis

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Time course observation of mouse skull development using microCT

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Modelling lung branching morphogenesis

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In vitro vasculogenesis model revisited: Measurement of VEGF diffusion

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Visualizing morphogen distribution in lumen

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Mechanism of lung branching morphogenesis

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Expression of proximodistal markers during chick lung development

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Expression of proximodistal markers during chick lung development

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Mechamism of lung branching morphogenesis

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Roles of epithelium and mesenchyme during lung branching morphogenesis

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Mechanism of lung branching morphogenesis

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Modeling lung branching morphogenesis via epithelial-mesenchymal interaction

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Modeling lung branching morphogenesis via epithelial-mesenchymal interaction

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How well does Turing's theory of morphogenesis work?

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Visualizing morphogen diffusion dynamics during lung branching morphogenesis in vitro.

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Visualizing Morphogen Diffusion Dynamics in lung branching morphogenesis

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A novel mathematical model of endothelial cell dynamics during angiogenic morphogenesis: analysis of cell-based mechanisms

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A New Mathematical Model of Pattern Formation of the Cranial Suture

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Modeling pattern formation of skull suture interdigitation

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Mathematical model of epithelial buckling for single- and multi-step processes of the intestinal villus formation

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Modeling vascular scaling law formation in retina vasculature

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Modeling vascular scaling law formation in retina vasculature

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Mechanism of jigsaw-puzzle pattern formation in plant leaf epidermal cell

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Analyses of sizes of self-organized patterns composed by endothelial cell

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記述言語：日本語  

記述言語：日本語  

記述言語：日本語  

記述言語：日本語  

記述言語：日本語  

灌流可能な血管を有するオンチップ腫瘍モデルの創出

記述言語：日本語  

3D血管網を用いた剪断応力の知覚と伝達による血管新生評価システム

記述言語：英語  

Remodeling in synthetic vascular network - experiment and modeling

DOI： 10.1016/j.mod.2017.04.059

記述言語：英語  

Development of three-dimensional tumor model with a perfusable vasculature using a microfluidic device

DOI： 10.1016/j.mod.2017.04.041

記述言語：英語  

Reconstitutive analyses of impacts of pericytes and blood flow on angiogenic morphogenesis using a microfluidic device

DOI： 10.1016/j.mod.2017.04.158

記述言語：日本語  

Anastomosis between tissue models and microchannels using angiogenic sprouts for the perfusable culture system

記述言語：日本語  

記述言語：日本語  

記述言語：日本語   掲載種別：記事・総説・解説・論説等（学術雑誌）  

生物の発生段階において、自発的なパターン形成によって周期構造が生じる事がある。数理生物学の分野ではこの現象をモデル化する際に Turing の反応拡散系が広く使われているが、発生生物学での応用例はま だ少ない。我々は四肢の骨格のパターン形成に関して Turing 系を応用 する試みをしているが、これによって実験的に進歩した部分、理論的に 進歩した部分をそれぞれ紹介し、数理生物学との相互作用による利点や 将来の展望について論じる。

リポジトリ公開URL： http://hdl.handle.net/2324/4751312

種別：査読等 

外国語雑誌　査読論文数：7

種別：査読等 

外国語雑誌　査読論文数：11

種別：査読等 

外国語雑誌　査読論文数：8

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世界初、植物細胞の形の謎を解明 鍵は頭蓋骨縫合線と数学 九州大学ほか

Turing vs. Wolpert - 反応拡散系に寄るパターン形成の応用研究の系譜

サイエンスZERO シリーズ細胞の世界１ 体をつくる不思議な"波”～

シリーズ細胞の世界（１）体を作る不思議な「波」

動物たちの模様を解く魔法の数式 天才数学者が残した遺産