Terminus-Selective Covalent Immobilization of Heparin on a Thermoresponsive Surface Using Click Chemistry for Efficient Binding of Basic Fibroblast Growth Factor.

Cell therapy using endothelial cells (ECs) has great potential for the treatment of congenital disorders, such as hemophilia A. Cell sheet technology utilizing a thermoresponsive culture dish is a promising approach to efficiently transplant donor cells. In this study, a new method to prepare terminus-selective heparin-immobilized thermoresponsive culture surfaces is developed to facilitate the preparation of EC sheets. Alkynes are introduced to the reducing terminus of heparin via reductive amination. Cu-catalyzed azide-alkyne cycloaddition (CuAAC) facilitates efficient immobilization of the terminus of heparin on a thermoresponsive surface, resulting in a higher amount of immobilized heparin while preserving its function. Heparin-immobilized thermoresponsive surfaces prepared using CuAAC exhibit good adhesion to human endothelial colony-forming cells (ECFCs). In addition, upon further binding to basic fibroblast growth factor (bFGF) on heparin-immobilized surfaces, increased proliferation of ECFCs on the surface is observed. The confluent ECFC monolayer cultured on bFGF-bound heparin-immobilized thermoresponsive surfaces exhibits relatively high fibronectin accumulation and cell number and detaches at 22 °C while maintaining the sheet-like structure. Because heparin has an affinity for several types of bioactive molecules, the proposed method can be applied to facilitate efficient cultures and sheet formations of various cell types.

Observation of the Anisotropic Magneto-Thomson Effect.

We report the observation of the anisotropic magneto-Thomson effect (AMTE), which is one of the higher-order thermoelectric effects in a ferromagnet. Using lock-in thermography, we demonstrated that in a ferromagnetic NiPt alloy, the cooling or heating induced by the Thomson effect depends on the angle between the magnetization direction and the temperature gradient or charge current applied to the alloy. AMTE observed here is the missing ferromagnetic analog of the magneto-Thomson effect in a nonmagnetic conductor, providing the basis for nonlinear spin caloritronics and thermoelectrics.

Regulation of von Willebrand factor by ADAMTS13 ameliorates lipopolysaccharide-induced lung injury in mice.

The relationship between von Willebrand factor (VWF) and inflammation has attracted considerable attention in recent years. VWF, which is stored in the Weibel-Palade bodies (WPBs) of endothelial cells (ECs), is released from WPBs in response to inflammatory stimuli and is thought to contribute to inflammation by promoting leukocyte extravasation. In this study, lung injury model mice were produced by intratracheal injection with lipopolysaccharides. The severity of lung inflammation was evaluated in mice with different genotypes (wild-type, Vwf-/-, Adamts13-/-) and mice treated with drugs that inhibit VWF function. Lung inflammation was significantly ameliorated in Vwf-/- mice compared with wild-type mice. Furthermore, inflammation was significantly suppressed in wild-type mice treated with anti-VWF A1 antibody or recombinant human ADAMTS13 compared with the untreated control group. The underlying mechanism appears to be an increased VWF/ADAMTS13 ratio at the site of inflammation and the interaction between blood cell components, such as leukocytes and platelets, and the VWF A1 domain, which promotes leukocyte infiltration into the lung. This study suggested that ADAMTS13 protein and other VWF-targeting agents may be a novel therapeutic option for treatment of pulmonary inflammatory diseases.

Generation of functional liver sinusoidal endothelial-like cells from human bone marrow-derived mesenchymal stem cells.

Introduction: Liver sinusoidal endothelial cells (LSECs) are specialized vascular endothelial cells that play an important role in the maintenance of biological homeostasis. However, the lack of versatile human LSECs has hindered research on LSECs and development of medical technologies for liver diseases including hemophilia A. In this study, we developed a technique to induce LSEC differentiation from human bone marrow-derived mesenchymal stem cells (BM-MSCs). Methods: To induce LSECs from human BM-MSCs, cytokines and chemical compounds associated with signaling implicated in LSEC differentiation and liver development were screened. Then LSEC-related genes and proteins expression in the differentiated cells were analyzed by qPCR and flow cytometry analysis, respectively. LSEC-related functions of the differentiated cells were also examined. Results: We found that the gene expression of LSEC markers, such as LYVE1, was considerably increased by culturing human BM-MSCs with bone morphogenetic protein 4, fibroblast growth factor 8b, transforming growth factor-ß signal inhibitor, and cyclic AMP. Furthermore, the differentiated cells expressed LSEC marker proteins and clearly demonstrated LSEC-specific functions, such as the uptake of hyaluronic acid. Conclusions: Our result indicate that the functional LSEC-like cells were successfully generated from human BM-MSCs using our established protocol.

Development of a method of passaging and freezing human iPS cell-derived hepatocytes to improve their functions.

Human induced pluripotent stem (iPS) cell-derived hepatocyte-like cells (HLCs) are expected to replace primary human hepatocytes as a new source of functional hepatocytes in various medical applications. However, the hepatic functions of HLCs are still low and it takes a long time to differentiate them from human iPS cells. Furthermore, HLCs have very low proliferative capacity and are difficult to be passaged due to loss of hepatic functions after reseeding. To overcome these problems, we attempted to develop a technology to dissociate, cryopreserve, and reseed HLCs in this study. By adding epithelial-mesenchymal transition inhibitors and optimizing the cell dissociation time, we have developed a method for passaging HLCs without loss of their functions. After passage, HLCs showed a hepatocyte-like polygonal cell morphology and expressed major hepatocyte marker proteins such as albumin and cytochrome P450 3A4 (CYP3A4). In addition, the HLCs had low-density lipoprotein uptake and glycogen storage capacity. The HLCs also showed higher CYP3A4 activity and increased gene expression levels of major hepatocyte markers after passage compared to before passage. Finally, they maintained their functions even after their cryopreservation and re-culture. By applying this technology, it will be possible to provide ready-to-use availability of cryopreserved HLCs for drug discovery research.

Charge-to-spin conversion in fully epitaxial Ru/Cu hybrid nanolayers with interface control.

The demonstration of the charge-to-spin conversion, especially with enhanced spin Hall conductivity, is crucial for the development of energy-efficient spintronic devices such as spin-orbit torque (SOT) based magnetoresistive random access memories. In this work, fully epitaxial Ru/Cu heterostructures were fabricated with interface engineering and nanolayer insertions consisting of Cu (1 nm)/Ru (1 nm) structures with different numbers of periods. The atomically controlled interface was confirmed by the high-resolution high-angle annular dark-field scanning transmission electron microscopy, and the epitaxial relationship persists even in the hybrid nanolayer insertion structures. The spin current generation was detected by the measurement of unidirectional spin Hall magnetoresistance, and the effective damping-like spin Hall efficiency (ξDL) was further quantitatively evaluated by the spin-torque ferromagnetic resonance with thickness dependence of the ferromagnetic layer. It is found that the sharp interface Ru/Cu film has a sizeableξDLof -2.2% and the insertion of Cu/Ru nanolayers at the interface can increase theξDLvalue to -3.7%. The former could be attributed to the interface spin-orbit filtering effect and the latter may be further understood by the intrinsic contribution from the local electronic structure tuning due to the lattice distortion near the interface. A large effective spin Hall conductivity is achieved to be (3∼5) × 105ℏ2eΩ-1m-1in the epitaxial Ru/Cu hybrid nanolayers, which is in the same range as that of platinum. This work indicates that the interfacial control with hybrid nanolayer structures can extend the SOT-based materials to highly conductive metals, even with weak spin-orbit interactions, toward high stability, low cost, and low energy consumption for spintronic applications.

Spin-orbit torque driven magnetization switching in W/CoFeB/MgO-based type-Y three terminal magnetic tunnel junctions.

We have studied current induced magnetization switching in W/CoFeB/MgO based three terminal magnetic tunnel junctions. The switching driven by spin-orbit torque (SOT) is evaluated in the so-called type-Y structure, in which the magnetic easy-axis of the CoFeB layer lies in the film plane and is orthogonal to the current flow. The effective spin Hall angle estimated from the bias field dependence of critical current (Ic) is ~ 0.07. The field and current dependence of the switching probability are studied. The field and DC current induced switching can be described using a model based on thermally assisted magnetization switching. In contrast, the 50 ns long pulse current dependence of the switching probability shows significant deviation from the model, even if contribution from the field-like torque is included. The deviation is particularly evident when the threshold switching current is larger. These results show that conventional thermally assisted magnetization switching model cannot be used to describe SOT induced switching using short current pulses.

Heusler alloys for spintronic devices: review on recent development and future perspectives.

Heusler alloys are theoretically predicted to become half-metals at room temperature (RT). The advantages of using these alloys are good lattice matching with major substrates, high Curie temperature above RT and intermetallic controllability for spin density of states at the Fermi energy level. The alloys are categorised into half- and full-Heusler alloys depending upon the crystalline structures, each being discussed both experimentally and theoretically. Fundamental properties of ferromagnetic Heusler alloys are described. Both structural and magnetic characterisations on an atomic scale are typically carried out in order to prove the half-metallicity at RT. Atomic ordering in the films is directly observed by X-ray diffraction and is also indirectly probed via the temperature dependence of electrical resistivity. Element specific magnetic moments and spin polarisation of the Heusler alloy films are directly measured using X-ray magnetic circular dichroism and Andreev reflection, respectively. By employing these ferromagnetic alloy films in a spintronic device, efficient spin injection into a non-magnetic material and large magnetoresistance are also discussed. Fundamental properties of antiferromagnetic Heusler alloys are then described. Both structural and magnetic characterisations on an atomic scale are shown. Atomic ordering in the Heusler alloy films is indirectly measured by the temperature dependence of electrical resistivity. Antiferromagnetic configurations are directly imaged by X-ray magnetic linear dichroism and polarised neutron reflection. The applications of the antiferromagnetic Heusler alloy films are also explained. The other non-magnetic Heusler alloys are listed. A brief summary is provided at the end of this review.

[Generation of Zone-specific Hepatocyte-like Cells from Human Induced Pluripotent Stem Cells for Accurate Prediction of Drug-induced Hepatotoxicity].

Human induced pluripotent stem (iPS) cell-derived hepatocyte-like cells (iPS-HLCs) are expected to be applicable to large-scale in vitro hepatotoxicity screening systems. Accordingly, methods for generating HLCs from human iPS cells have been improved over the past decade. However, although human hepatocytes have zone-specific characteristics in vivo, there is currently no technique to generate zone-specific HLCs from human iPS cells. Therefore, to generate HLCs with zone-specific properties from human iPS cells, we cultured iPS-HLCs using a parenchymal or nonparenchymal cell-conditioned medium (CM). The results showed that urea production and gluconeogenesis capacity in iPS-HLCs were increased by culturing with cholangiocyte-CM, and glutamine production and drug metabolism capacity in iPS-HLCs were increased by culturing with hepatocyte-CM. It was thus clarified that iPS-HLCs acquire zone 1 hepatocyte-like properties by culturing with cholangiocyte-CM and that iPS-HLCs acquire zone 3 hepatocyte-like properties by culturing with hepatocyte-CM. In addition, we found that WNT inhibitory factor-1 secreted from cholangiocytes, and WNT7B and WNT8B secreted from hepatocytes play important roles in the zone-specific conversion of iPS-HLCs. We hope that our findings will facilitate the application of iPS-HLCs to drug discovery research.

Erratum: Realizing Room-Temperature Resonant Tunnel Magnetoresistance in Cr/Fe/MgAl2O4 Quasi-Quantum Well Structures.

[This corrects the article DOI: 10.1002/advs.201901438.].

Realizing Room-Temperature Resonant Tunnel Magnetoresistance in Cr/Fe/MgAl2O4 Quasi-Quantum Well Structures.

The quantum well (QW) realizes new functionalities due to the discrete electronic energy levels formed in the well-shaped potential. Magnetic tunnel junctions (MTJs) combined with a quasi-QW structure of Cr/ultrathin-Fe/MgAl2O4(001)/Fe, in which the Cr quasi-barrier layer confines Δ 1 up-spin electrons to the Fe well, are prepared with perfectly lattice-matched interfaces and atomic layer number control. Resonant peaks are clearly observed in the differential conductance of the MTJs due to the formation of QWs. Furthermore, enhanced tunnel magnetoresistance (TMR) peaks at the resonant bias voltages are realized for the MTJs at room temperature, i.e., it is observed that TMR ratios at specific and even high bias-voltages (V bias) are larger than zero-bias TMR ratios for the MTJs with odd Fe atomic layers, in contrast to the earlier experimental studies. In addition, a new finding in this study is unique sign changes in the temperature coefficient of resistance (TCR) depending on the Fe thickness and V bias, which is interpreted as a signature of the QW formation of Δ1 symmetry electronic states. The present study suggests that the spin-dependent resonant tunneling via the QWs formed in Cr/ultrathin-Fe/MgAl2O4/Fe structures should open a new pathway to achieve a large TMR at practically high V bias.

The spin Nernst effect in tungsten.

The spin Hall effect allows the generation of spin current when charge current is passed along materials with large spin-orbit coupling. It has been recently predicted that heat current in a nonmagnetic metal can be converted into spin current via a process referred to as the spin Nernst effect. We report the observation of the spin Nernst effect in W. In W/CoFeB/MgO heterostructures, we find changes in the longitudinal and transverse voltages with magnetic field when temperature gradient is applied across the film. The field dependence of the voltage resembles that of the spin Hall magnetoresistance. A comparison of the temperature gradient-induced voltage and the spin Hall magnetoresistance allows direct estimation of the spin Nernst angle. We find the spin Nernst angle of W to be similar in magnitude but opposite in sign to its spin Hall angle. Under an open-circuit condition, this sign difference results in the spin current generation larger than otherwise. These results highlight the distinct characteristics of the spin Nernst and spin Hall effects, providing pathways to explore materials with unique band structures that may generate large spin current with high efficiency.

Human ESC/iPSC-Derived Hepatocyte-like Cells Achieve Zone-Specific Hepatic Properties by Modulation of WNT Signaling.

The function of hepatocytes largely depends on their position in the liver lobule. Although the method of differentiating hepatocytes from human pluripotent stem cells has been largely improved over the past decade, there remains no technique for generating hepatocyte-like cells (HLCs) with zone-specific hepatic properties. In this study, we searched for the factors that promote acquisition of zone-specific properties of HLCs. Here, we identified that WNT7B and WNT8B secreted from hepatocytes and cholangiocytes play important roles in achieving perivenous zone-specific characteristics, such as the enhancement of glutamine secretion, citric acid cycle, cytochrome P450 (CYP) 1A2 metabolism, and CYP1A2 induction capacities. We also found that WNT inhibitory factor (WIF-1) secreted from cholangiocytes was necessary for achieving periportal zone-specific characteristics, such as the enhancement of urea secretion and gluconeogenesis capacities. Therefore, WNT signal modulators secreted from hepatocytes or cholangiocytes conferred zone-specific hepatic properties onto HLCs.

Human induced-pluripotent stem cell-derived hepatocyte-like cells as an in vitro model of human hepatitis B virus infection.

In order to understand the life cycle of hepatitis B virus (HBV) and to develop efficient anti-HBV drugs, a useful in vitro cell culture system which allows HBV infection and recapitulates virus-host interactions is essential; however, pre-existing in vitro HBV infection models are often problematic. Here, we examined the potential of human induced-pluripotent stem (iPS) cell-derived hepatocyte-like cells (iPS-HLCs) as an in vitro HBV infection model. Expression levels of several genes involved in HBV infection, including the sodium taurocholate cotransporting polypeptide (NTCP) gene, were gradually elevated as the differentiation status of human iPS cells proceeded to iPS-HLCs. The mRNA levels of these genes were comparable between primary human hepatocytes (PHHs) and iPS-HLCs. Following inoculation with HBV, we found significant production of HBV proteins and viral RNAs in iPS-HLCs. The three major forms of the HBV genome were detected in iPS-HLCs by Southern blotting analysis. Anti-HBV agents entecavir and Myrcludex-B, which are a nucleoside analogue reverse transcriptase inhibitor and a synthetic pre-S1 peptide, respectively, significantly inhibited HBV infection in iPS-HLCs. These data demonstrate that iPS-HLCs can be used as a promising in vitro HBV infection model.

Voltage control of magnetic anisotropy in epitaxial Ru/Co2FeAl/MgO heterostructures.

Voltage control of magnetic anisotropy (VCMA) in magnetic heterostructures is a key technology for achieving energy-efficiency electronic devices with ultralow power consumption. Here, we report the first demonstration of the VCMA effect in novel epitaxial Ru/Co2FeAl(CFA)/MgO heterostructures with interfacial perpendicular magnetic anisotropy (PMA). Perpendicularly magnetized tunnel junctions with the structure of Ru/CFA/MgO were fabricated and exhibited an effective voltage control on switching fields for the CFA free layer. Large VCMA coefficients of 108 and 139 fJ/Vm for the CFA film were achieved at room temperature and 4 K, respectively. The interfacial stability in the heterostructure was confirmed by repeating measurements. Temperature dependences of both the interfacial PMA and the VCMA effect were also investigated. It is found that the temperature dependences follow power laws of the saturation magnetization with an exponent of ~2, where the latter is definitely weaker than that of conventional Ta/CoFeB/MgO. The significant VCMA effect observed in this work indicates that the Ru/CFA/MgO heterostructure could be one of the promising candidates for spintronic devices with voltage control.

Degradation Characteristics of MgO Based Magnetic Tunnel Junction Caused by Surface Roughness of Ta/Ru Buffer Layers.

We investigated how surface roughness of a Ta/Ru buffer layer affects the degradation characteristics on MgO-based magnetic tunnel junctions (MTJs). MTJs with worse surface roughness on the buffer layer showed increased resistance drift and degraded time-dependent dielectric breakdown (TDDB) characteristics. We suggest that this resulted from reduced MgO thickness on the MTJ with worse surface roughness on the buffer layer, which was estimated by the TDDB and analytic approach. As a result, surface roughness of the buffer layer is a critical factors that impacts the reliability of MTJs, and it should be controlled to have the smallest roughness value as possible.

Laminin 411 and 511 promote the cholangiocyte differentiation of human induced pluripotent stem cells.

The drug discovery research for cholestatic liver diseases has been hampered by the lack of a well-established human cholangiocyte model. Functional cholangiocyte-like cells differentiated from human induced pluripotent stem (iPS) cells are expected to be a promising candidate for such research, but there remains no well-established method for differentiating cholangiocytes from human iPS cells. In this study, we searched for a suitable extracellular matrix to promote cholangiocyte differentiation from human iPS cells, and found that both laminin 411 and laminin 511 were suitable for this purpose. The gene expression levels of the cholangiocyte markers, aquaporin 1 (AQP1), SRY-box 9 (SOX9), cystic fibrosis transmembrane conductance regulator (CFTR), G protein-coupled bile acid receptor 1 (GPBAR1), Jagged 1 (JAG1), secretin receptor (SCTR), and γ-glutamyl transferase (GGT1) were increased by using laminin 411 or laminin 511 as a matrix. In addition, the percentage of AQP1-positive cells was increased from 61.8% to 92.5% by using laminin 411 or laminin 511. Furthermore, the diameter and number of cysts consisted of cholangiocyte-like cells were increased when using either matrix. We believe that the human iPS cell-derived cholangiocyte-like cells, which were generated by using our differentiation technology, would be useful for the drug discovery research of cholestatic liver diseases.

Spin Hall Magnetoresistance in Metallic Bilayers.

Spin Hall magnetoresistance (SMR) is studied in metallic bilayers that consist of a heavy metal (HM) layer and a ferromagnetic metal (FM) layer. We find a nearly tenfold increase of SMR in W/CoFeB compared to previously studied HM/ferromagnetic insulator systems. The SMR increases with decreasing temperature despite the negligible change in the W layer resistivity. A model is developed to account for the absorption of the longitudinal spin current to the FM layer, one of the key characteristics of a metallic ferromagnet. We find that the model not only quantitatively describes the HM layer thickness dependence of SMR, allowing accurate estimation of the spin Hall angle and the spin diffusion length of the HM layer, but also can account for the temperature dependence of SMR by assuming a temperature dependent spin polarization of the FM layer. These results illustrate the unique role a metallic ferromagnetic layer plays in defining spin transmission across the HM/FM interface.

Hepatic maturation of human iPS cell-derived hepatocyte-like cells by ATF5, c/EBPα, and PROX1 transduction.

Hepatocyte-like cells differentiated from human iPS cells (human iPS-HLCs) are expected to be utilized in drug development and research. However, recent hepatic characterization of human iPS-HLCs showed that these cells resemble fetal hepatocytes rather than adult hepatocytes. Therefore, in this study, we aimed to develop a method to enhance the hepatic function of human iPS-HLCs. Because the gene expression levels of the hepatic transcription factors (activating transcription factor 5 (ATF5), CCAAT/enhancer-binding protein alpha (c/EBPα), and prospero homeobox protein 1 (PROX1)) in adult liver were significantly higher than those in human iPS-HLCs and fetal liver, we expected that the hepatic functions of human iPS-HLCs could be enhanced by adenovirus (Ad) vector-mediated ATF5, c/EBPα, and PROX1 transduction. The gene expression levels of cytochrome P450 (CYP) 2C9, 2E1, alpha-1 antitrypsin, transthyretin, Na+/taurocholate cotransporting polypeptide, and uridine diphosphate glucuronosyl transferase 1A1 and protein expression levels of CYP2C9 and CYP2E1 were upregulated by ATF5, c/EBPα, and PROX1 transduction. These results suggest that the hepatic functions of the human iPS-HLCs could be enhanced by ATF5, c/EBPα, and PROX1 transduction. Our findings would be useful for the hepatic maturation of human iPS-HLCs.

Enhanced orbital magnetic moments in magnetic heterostructures with interface perpendicular magnetic anisotropy.

We have studied the magnetic layer thickness dependence of the orbital magnetic moment in magnetic heterostructures to identify contributions from interfaces. Three different heterostructures, Ta/CoFeB/MgO, Pt/Co/AlOx and Pt/Co/Pt, which possess significant interface contribution to the perpendicular magnetic anisotropy, are studied as model systems. X-ray magnetic circular dichroism spectroscopy is used to evaluate the relative orbital moment, i.e. the ratio of the orbital to spin moments, of the magnetic elements constituting the heterostructures. We find that the relative orbital moment of Co in Pt/Co/Pt remains constant against its thickness whereas the moment increases with decreasing Co layer thickness for Pt/Co/AlOx, suggesting that a non-zero interface orbital moment exists for the latter system. For Ta/CoFeB/MgO, a non-zero interface orbital moment is found only for Fe. X-ray absorption spectra shows that a particular oxidized Co state in Pt/Co/AlOx, absent in other heterosturctures, may give rise to the interface orbital moment in this system. These results show element specific contributions to the interface orbital magnetic moments in ultrathin magnetic heterostructures.