Identification and removal of unexpected proliferative off-target cells emerging after iPSC-derived pancreatic islet cell implantation.

Differentiation of pancreatic endocrine cells from human pluripotent stem cells (PSCs) has been thoroughly investigated for application in cell therapy against diabetes. In the context of induced pancreatic endocrine cell implantation, previous studies have reported graft enlargement resulting from off-target pancreatic lineage cells. However, there is currently no documented evidence of proliferative off-target cells beyond the pancreatic lineage in existing studies. Here, we show that the implantation of seven-stage induced PSC-derived pancreatic islet cells (s7-iPICs) leads to the emergence of unexpected off-target cells with proliferative capacity via in vivo maturation. These cells display characteristics of both mesenchymal stem cells (MSCs) and smooth muscle cells (SMCs), termed proliferative MSC- and SMC-like cells (PMSCs). The frequency of PMSC emergence was found to be high when 108 s7-iPICs were used. Given that clinical applications involve the use of a greater number of induced cells than 108, it is challenging to ensure the safety of clinical applications unless PMSCs are adequately addressed. Accordingly, we developed a detection system and removal methods for PMSCs. To detect PMSCs without implantation, we implemented a 4-wk-extended culture system and demonstrated that putative PMSCs could be reduced by compound treatment, particularly with the taxane docetaxel. When docetaxel-treated s7-iPICs were implanted, the PMSCs were no longer observed. This study provides useful insights into the identification and resolution of safety issues, which are particularly important in the field of cell-based medicine using PSCs.

Near-Infrared Emission from Tin-Lead (Sn-Pb) Alloyed Perovskite Quantum Dots by Sodium Doping.

Phase-stable CsSnx Pb1-x I3 perovskite quantum dots (QDs) hold great promise for optoelectronic applications owing to their strong response in the near-infrared region. Unfortunately, optimal utilization of their potential is limited by the severe photoluminescence (PL) quenching, leading to extremely low quantum yields (QYs) of approximately 0.3 %. The ultra-low sodium (Na) doping presented herein is found to be effective in improving PL QYs of these alloyed QDs without alerting their favourable electronic structure. X-ray photoelectron spectroscopy (XPS) studies suggest the formation of a stronger chemical interaction between I- and Sn2+ ions upon Na doping, which potentially helps to stabilize Sn2+ and suppresses the formation of I vacancy defects. The optimized PL QY of the Na-doped QDs reaches up to around 28 %, almost two orders of magnitude enhancement compared with the pristine one.

Highly Efficient 17.6% Tin-Lead Mixed Perovskite Solar Cells Realized through Spike Structure.

Frequently observed high Voc loss in tin-lead mixed perovskite solar cells is considered to be one of the serious bottle-necks in spite of the high attainable Jsc due to wide wavelength photon harvesting. An amicable solution to minimize the Voc loss up to 0.50 V has been demonstrated by introducing an n-type interface with spike structure between the absorber and electron transport layer inspired by highly efficient Cu(In,Ga)Se2 solar cells. Introduction of a conduction band offset of ∼0.15 eV with a thin phenyl-C61-butyric acid methyl ester layer (∼25 nm) on the top of perovskite absorber resulted into improved Voc of 0.75 V leading to best power conversion efficiency of 17.6%. This enhancement is attributed to the facile charge flow at the interface owing to the reduction of interfacial traps and carrier recombination with spike structure as evidenced by time-resolved photoluminescence, nanosecond transient absorption, and electrochemical impedance spectroscopy measurements.

Generation of branching ureteric bud tissues from human pluripotent stem cells.

Recent progress in kidney regeneration research is noteworthy. However, the selective and robust differentiation of the ureteric bud (UB), an embryonic renal progenitor, from human pluripotent stem cells (hPSCs) remains to be established. The present study aimed to establish a robust induction method for branching UB tissue from hPSCs towards the creation of renal disease models. Here, we found that anterior intermediate mesoderm (IM) differentiates from anterior primitive streak, which allowed us to successfully develop an efficient two-dimensional differentiation method of hPSCs into Wolffian duct (WD) cells. We also established a simplified procedure to generate three-dimensional WD epithelial structures that can form branching UB tissues. This system may contribute to hPSC-based regenerative therapies and disease models for intractable disorders arising in the kidney and lower urinary tract.

Identification of a small molecule that facilitates the differentiation of human iPSCs/ESCs and mouse embryonic pancreatic explants into pancreatic endocrine cells.

AIMS/HYPOTHESIS: Pancreatic beta-like cells generated from human induced pluripotent stem cells (hiPSCs) or human embryonic stem cells (hESCs) offer an appealing donor tissue source. However, differentiation protocols that mainly use growth factors are costly. Therefore, in this study, we aimed to establish efficient differentiation protocols to change hiPSCs/hESCs to insulin (INS)+ cells using novel small-molecule inducers. METHODS: We screened small molecules that increased the induction rate of INS+ cells from hESC-derived pancreatic and duodenal homeobox 1 (PDX1)+ pancreatic progenitor cells. The differentiation protocol to generate INS+ cells from hiPSCs/hESCs was optimised using hit compounds, and INS+ cells induced with the compounds were characterised for their in vitro and in vivo functions. The inducing activity of the hit compounds was also examined using mouse embryonic pancreatic tissues in an explant culture system. Finally, RNA sequencing analyses were performed on the INS+ cells to elucidate the mechanisms of action by which the hit compounds induced pancreatic endocrine differentiation. RESULTS: One hit compound, sodium cromoglicate (SCG), was identified out of approximately 1250 small molecules screened. When SCG was combined with a previously described protocol, the induction rate of INS+ cells increased from a mean ± SD of 5.9 ± 1.5% (n = 3) to 16.5 ± 2.1% (n = 3). SCG induced neurogenin 3-positive cells at a mean ± SD of 32.6 ± 4.6% (n = 3) compared with 14.2 ± 3.6% (n = 3) for control treatment without SCG, resulting in an increased generation of endocrine cells including insulin-producing cells. Similar induction by SCG was confirmed using mouse embryonic pancreatic explants. We also confirmed that the mechanisms of action by which SCG induced pancreatic endocrine differentiation included the inhibition of bone morphogenetic protein 4 signalling. CONCLUSIONS/INTERPRETATION: SCG improves the generation of pancreatic endocrine cells from multiple hiPSC/hESC lines and mouse embryonic pancreatic explants by facilitating the differentiation of endocrine precursors. This discovery will contribute to elucidating the mechanisms of pancreatic endocrine development and facilitate cost-effective generation of INS+ cells from hiPSCs/hESCs. DATA AVAILABILITY: The RNA sequencing data generated during the current study are available in the Gene Expression Omnibus ( www.ncbi.nlm.nih.gov/geo ) with series accession number GSE89973.

Colloidal Synthesis of Air-Stable Alloyed CsSn1-xPbxI3 Perovskite Nanocrystals for Use in Solar Cells.

Organic-inorganic hybrid perovskite solar cells have demonstrated unprecedented high power conversion efficiencies in the past few years. Now, the universal instability of the perovskites has become the main barrier for this kind of solar cells to realize commercialization. This situation can be even worse for those tin-based perovskites, especially for CsSnI3, because upon exposure to ambient atmosphere the desired black orthorhombic phase CsSnI3 would promptly lose single crystallinity and degrade to the inactive yellow phase, followed by irreversible oxidation into metallic Cs2SnI6. By alloying CsSnI3 with CsPbI3, we herein report the synthesis of alloyed perovskite quantum dot (QD), CsSn1-xPbxI3, which not only can be phase-stable for months in purified colloidal solution but also remains intact even directly exposed to ambient air, far superior to both of its parent CsSnI3 and CsPbI3 QDs. Ultrafast transient absorption spectroscopy studies reveal that the photoexcited electrons in the alloyed QDs can be injected into TiO2 nanocrystals at a fast rate of 1.12 × 1011 s-1, which enables a high photocurrent generation in solar cells.

Modelling urea-cycle disorder citrullinemia type 1 with disease-specific iPSCs.

Citrullinemia type 1 (CTLN1) is a urea cycle disorder (UCD) caused by mutations of the ASS1 gene, which is responsible for production of the enzyme argininosuccinate synthetase (ASS), and classically presented as life-threatening hyperammonemia in newborns. Therapeutic options are limited, and neurological sequelae may persist. To understand the pathophysiology and find novel treatments, induced pluripotent stem cells (iPSCs) were generated from a CTLN1 patient and differentiated into hepatocyte-like cells (HLCs). CTLN1-HLCs have lower ureagenesis, recapitulating part of the patient's phenotype. l-arginine, an amino acid clinically used for UCD treatment, improved this phenotype in vitro. Metabolome analysis revealed an increase in tricarboxylic acid (TCA) cycle metabolites in CTLN1, suggesting a connection between CTLN1 and the TCA cycle. This CTLN1-iPSC model improves the understanding of CTLN1 pathophysiology and can be used to pursue new therapeutic approaches.

Ligand-dependent exciton dynamics and photovoltaic properties of PbS quantum dot heterojunction solar cells.

The surface chemistry of colloidal quantum dots (QDs) plays an important role in determining the photoelectric properties of QD films and the corresponding quantum dot heterojunction solar cells (QDHSCs). To investigate the effects of the ligand structure on the photovoltaic performance and exciton dynamics of QDHSCs, PbS QDHSCs were fabricated by the solid state ligand exchange method with mercaptoalkanoic acid as the cross-linking ligand. Temperature-dependent photoluminescence and ultrafast transient absorption spectra show that the electronic coupling and charge transfer rate within QD ensembles were monotonically enhanced as the ligand length decreased. However, in practical QDHSCs, the second shortest ligand 3-mercaptopropionic acid (MPA) showed higher power conversion efficiency than the shortest ligand thioglycolic acid (TGA). This could be attributed to the difference in their surface trap states, supported by thermally stimulated current measurements. Moreover, compared with the non-conjugated ligand MPA, the conjugated ligand 4-mercaptobenzoic acid (MBA) introduces less trap states and has a similar charge transfer rate in QD ensembles, but has poor photovoltaic properties. This unexpected result could be contributed by the QD-ligand orbital mixing, leading to the charge transfer from QDs to ligands instead of charge transfer between adjacent QDs. This work highlights the significant effects of ligand structures on the photovoltaic properties and exciton dynamics of QDHSCs, which would shed light on the further development of QD-based photoelectric devices.

Redefining definitive endoderm subtypes by robust induction of human induced pluripotent stem cells.

Many reports have described methods that induce definitive endoderm (DE) cells from human pluripotent stem cells (hPSCs). However, it is unclear whether the differentiation propensity of these DE cells is uniform. This uncertainty is due to the different developmental stages that give rise to anterior and posterior DE from anterior primitive streak (APS). Therefore, these DE cell populations might be generated from the different stages of APS cells, which affect the DE cell differentiation potential. Here, we succeeded in selectively differentiating early and late APS cells from human induced pluripotent stem cells (hiPSCs) using different concentrations of CHIR99021, a small molecule Wnt/ß-catenin pathway activator. We also established novel differentiation systems from hiPSCs into three types of DE cells: anterior and posterior domains of anterior DE cells through early APS cells and posterior DE cells through late APS cells. These different DE cell populations could differentiate into distinct endodermal lineages in vitro, such as lung, liver or small intestine progenitors. These results indicate that different APS cells can produce distinct types of DE cells that have proper developmental potency and suggest a method to evaluate the quality of endodermal cell induction from hPSCs.

The cause for the low efficiency of dye sensitized solar cells with a combination of ruthenium dyes and cobalt redox.

It has been a concern that the cobalt redox cannot give a good performance for the dye-sensitized solar cells when it is used with ruthenium dyes. The electron dynamics measurements clarified the electron loss processes, and clarified the cause. The result indicated the direct interaction between the ruthenium dyes with the cobalt redox, and it reduced the charge injection from the triplet state of the dyes to the titanium oxide, and also it increased the electron recombination process with the cobalt redox species. Both the problems of injection and recombination were solved by using the ruthenium dye with alkyl chains keeping a distance between the dye and the cobalt redox.

Overexpression of TRB3 in muscle alters muscle fiber type and improves exercise capacity in mice.

Increasing evidence suggests that TRB3, a mammalian homolog of Drosophila tribbles, plays an important role in cell growth, differentiation, and metabolism. In the liver, TRB3 binds and inhibits Akt activity, whereas in adipocytes, TRB3 upregulates fatty acid oxidation. In cultured muscle cells, TRB3 has been identified as a potential regulator of insulin signaling. However, little is known about the function and regulation of TRB3 in skeletal muscle in vivo. In the current study, we found that 4 wk of voluntary wheel running (6.6 ± 0.4 km/day) increased TRB3 mRNA by 1.6-fold and protein by 2.5-fold in the triceps muscle. Consistent with this finding, muscle-specific transgenic mice that overexpress TRB3 (TG) had a pronounced increase in exercise capacity compared with wild-type (WT) littermates (TG: 1,535 ± 283; WT: 644 ± 67 joules). The increase in exercise capacity in TRB3 TG mice was not associated with changes in glucose uptake or glycogen levels; however, these mice displayed a dramatic shift toward a more oxidative/fatigue-resistant (type I/IIA) muscle fiber type, including threefold more type I fibers in soleus muscles. Skeletal muscle from TRB3 TG mice had significantly decreased PPARα expression, twofold higher levels of miR208b and miR499, and corresponding increases in the myosin heavy chain isoforms Myh7 and Myb7b, which encode these microRNAs. These findings suggest that TRB3 regulates muscle fiber type via a peroxisome proliferator-activated receptor-α (PPAR-α)-regulated miR499/miR208b pathway, revealing a novel function for TRB3 in the regulation of skeletal muscle fiber type and exercise capacity.

Control of charge dynamics through a charge-separation interface for all-solid perovskite-sensitized solar cells.

The relationship between the structure of the charge-separation interface and the photovoltaic performance of all-solid dye-sensitized solar cells is reported. This cell is composed of porous a TiO2/perovskite (CH3NH3PbI(x)Cl(3-x))/p-type organic conductor. The porous titania layer was passivated with Al2O3 or Y2O3 to remove surface traps of the porous titania layer. Both passivations were effective in increasing the efficiency of the solar cell. Especially, the effect of Y2O3 passivation was remarkable. After passivation, the efficiency increased from 6.59 to 7.5%. The increase in the efficiency was discussed in terms of the electron lifetime in TiO2, the thermally stimulated current, the measurement of the microwave refractive carrier lifetime, and transition absorption spectroscopy. It was proven that surface passivation resulted in retardation of charge recombination between the electrons in the porous titania layers and the holes in the p-type organic conductors.

Effect of electrolyte constituents on the motion of ionic species and recombination kinetics in dye-sensitized solar cells.

The dynamic motion of ions in electrolyte solutions and its effect on recombination was investigated by the heterodyne transient grating method in addition to transient absorption and transient photocurrent methods in dye sensitized solar cells. Realignment of ionic species at the electrode/electrolyte interface was observed after the electron injection in TiO2 on the order of µs. The process was affected by the total quantity of ionic species as well as cation species in the electrolyte. The recombination processes of the electrons were also affected by the constituents; the probability of the electron-electrolyte recombination decreased with decrease in I2 concentration; the dominant recombination process changed from the electron-electrolyte to the electron-dye recombination by decreasing I(-) concentration. It is concluded that sufficient I(-) is necessary for the suppression of the electron-dye recombination and that sufficient I2 is necessary for an efficient redox cycle, while low concentration of I3(-) ions at the electrolyte/TiO2 interface is preferable to suppress the electron-electrolyte recombination. The effect of the cation size in an electrolyte solution on the charge dynamics was also investigated, and it was revealed that the steric hindrance of cations changed the penetration of ionic species into the nanoporous dye/TiO2 electrode, causing a change in the electrostatic properties at the interface. The cation dependence indicated that the presence of large-sized cations suppressed the electron-electrolyte recombination by disturbing the approach of I3(-) paired with the cations.

Charge transfer and recombination at the metal oxide/CH3NH3PbClI2/spiro-OMeTAD interfaces: uncovering the detailed mechanism behind high efficiency solar cells.

In recent years, organometal halide perovskite-based solid-state hybrid solar cells have attracted unexpected increasing interest because of their high efficiency (the record power conversion efficiency has been reported to be over 15%) and low fabrication cost. It has been accepted that the high efficiency was mainly attributed to the strong optical absorption (absorption coefficient: 15,000 cm(-1) at 550 nm) over a broader range (up to 800 nm) and the long lifetimes of photoexcited charge carriers (in the order of 10 ns - a few 100 ns) of the perovskite absorbers. However, much of the fundamental photophysical properties of perovskite relating to the high photovoltaic performance are remained to be investigated. The charge separation and recombination processes at the material interfaces are particularly important for solar cell performances. To better understand the high efficiency of perovskite solar cells, we systematically investigated the charge separation (electron and hole injection) and charge recombination dynamics of CH3NH3PbClI2 hybrid solar cells employing TiO2 nanostructures as the electron transfer material (ETM) and spiro-OMeTAD as the hole transfer material (HTM). The measurements were carried out using transient absorption (TA) techniques on a time scale from sub-picoseconds to milliseconds. We clarified the timescales of electron injection, hole injection, and recombination processes in TiO2/CH3NH3PbClI2/spiro-OMeTAD solar cells. Charge separation and collection efficiency of the perovskite-based solar cells were discussed. In addition, the effect of TiO2 size on the charge separation and recombination dynamics was also investigated. It was found that all TiO2-based perovskite solar cells possessed similar charge separation processes, but quite different recombination dynamics. Our results indicate that charge recombination was crucial to the performance of the perovskite solar cells, which could be effectively suppressed through optimising nanostructured TiO2 films and surface passivation, thus pushing these cells to even higher efficiency.

Multiple exciton generation in cluster-free alloy Cd(x)Hg(1-x)Te colloidal quantum dots synthesized in water.

A number of different composition CdxHg1-xTe alloy quantum dots have been synthesized using a modified aqueous synthesis and ion exchange method. The benefits of good stoichiometric control and high emission quantum yield were retained whilst also ensuring that the tendency to form gel-like clusters and adsorb excess cations in the stabilizing ligand shells was mitigated using a sequestering method to remove excess ionic material during and after the synthesis. This was highly desirable for ultrafast carrier dynamics measurements, avoiding strong photocharging effects which may mask fundamental carrier signals. Transient grating measurements revealed a composition dependent carrier multiplication process which competes with phonon mediated carrier cooling to deplete the initial hot carrier population. The interplay between these two mechanisms is strongly dependent on the electron effective mass which in these alloys has a marked composition dependence and may be considerably lower than the hole effective mass. For a composition x = 0.52 we measured a maximum carrier multiplication quantum yield of 199 ± 19% with pump photon energy 3 times the bandgap energy, Eg, whilst the threshold energy is calculated to be just 2.15Eg. There is some evidence to suggest an impact ionization process analogous to the inverse Auger S mechanism seen in bulk CdxHg1-xTe.

Photoexcited Carrier Dynamics in Iodine-Doped CH3NH3PbBr3 Single Crystals.

Iodine-doped bromide perovskite single crystals (IBPSCs) have important applications in optoelectronic fields, such as in solar cells. Currently, much research has aimed to study the phase separation phenomenon and device performance improvements in IBPSCs. However, important intrinsic photoexcited carrier dynamics are often overlooked in IBPSCs. Here, we explored the photoexcited carrier dynamics in typical iodine-doped MAPbBr3 single crystals using the excitation intensity-dependent steady-state photoluminescence (PL) and time-resolved photoluminescence (TRPL) technique. We found that the trap state density changes with an increase in the amount of doped iodine. Further, we noticed that there is an influence of carrier diffusion on the photoexcited carrier dynamics, and then, we evaluated the carrier diffusion coefficients and recombination constants via numerical simulations of the PL kinetics. Consequently, we found that the electron shallow trap-related carrier behaviors substantially impacted the PL kinetics. Our results greatly facilitate a deeper understanding of the fundamental characteristics of mixed halide perovskite material.

Detection of non-absorbing charge dynamics via refractive index change in dye-sensitized solar cells.

The carrier dynamics in dye-sensitized solar cells was investigated by using the transient grating, in addition to the transient absorption method and transient photocurrent method on the order of microseconds to seconds. The signals for the same sample were obtained under a short-circuit condition to compare the carrier dynamics via refractive index change with the transient photocurrent measurement. Optically silent carrier dynamics by transient absorption have been successfully observed via a refractive index change. The corresponding signal components were originated from the charge dynamics at the solid/liquid interface, especially on the liquid side; rearrangement or diffusion motion of charged redox species occurred when the injected electrons were trapped at the TiO2 surface and when the electron-electrolyte recombination occurred at the interface. The assignments were confirmed from the dependence on the viscosity of the solvent and the presence of 4-tert-butyl pyridine. As the viscosity of the solvent increased, the rearrangement and the motion of the charged redox species were delayed. Since the rearrangement dynamics was changed by the presence of 4-tert-butyl pyridine, it affected not only the TiO2 surface but also the redox species close to the interface.

Carrier dynamics in quantum-dot sensitized solar cells measured by transient grating and transient absorption methods.

Carrier dynamics in quantum-dot sensitized solar cells (QDSSCs) was clarified by combining the information obtained by the heterodyne transient grating (HD-TG), transient absorption (TA) and transient photocurrent (TP) measurements under the short circuit conditions in the time range from microseconds to seconds. The HD-TG signal is sensitive to the ionic species at the electrode/electrolyte interface, and the electrons in the titanium oxide layer injected from quantum dots (QDs) were monitored by the TA signal, and the photocurrent as a final output was monitored by the TP signal. By using the compensating information, the whole picture of the charge dynamics was obtained in the time region after the initial electron injection from QDs into the titanium oxide layer. In the former part of this paper, the assignment of the responses for each measurement was clarified based on the previous paper on dye sensitized solar cells (S. Kuwahara, et al. Phys. Chem. Chem. Phys., 2013, 15(16), 5975-5981). In the latter part, the effect of the device parameters for actual QDSSCs, such as electrolyte concentrations, and coating times of surface passivation of QDs were investigated.

Huge suppression of charge recombination in P3HT-ZnO organic-inorganic hybrid solar cells by locating dyes at the ZnO/P3HT interfaces.

The charge separation and charge recombination dynamics in P3HT-ZnO and P3HT-dye-ZnO bulk heterojunction organic-inorganic hybrid solar cells (OIHSCs) prepared by a one-pot method were studied using a transient absorption (TA) method, both for optical absorption of P3HT in the visible region and for optical absorption of SQ36 in the NIR region. In the case of P3HT-ZnO, the charge separation was very fast, occurring within 1 ps. On the other hand, high charge recombination between electrons in the surface states and/or the conduction band of ZnO and holes in P3HT was observed. In the case of P3HT-dye-ZnO, we found that the charge recombination could be greatly suppressed by locating the dye at the P3HT/ZnO interfaces while maintaining a fast charge separation rate (a few ps to 10 ps). Our findings provide one methodology for the design of OIHSCs for improving their conversion efficiency, which is to position the dye at the appropriate BHJ interfaces.

Sucrose nonfermenting AMPK-related kinase (SNARK) mediates contraction-stimulated glucose transport in mouse skeletal muscle.

The signaling mechanisms that mediate the important effects of contraction to increase glucose transport in skeletal muscle are not well understood, but are known to occur through an insulin-independent mechanism. Muscle-specific knockout of LKB1, an upstream kinase for AMPK and AMPK-related protein kinases, significantly inhibited contraction-stimulated glucose transport. This finding, in conjunction with previous studies of ablated AMPKalpha2 activity showing no effect on contraction-stimulated glucose transport, suggests that one or more AMPK-related protein kinases are important for this process. Muscle contraction increased sucrose nonfermenting AMPK-related kinase (SNARK) activity, an effect blunted in the muscle-specific LKB1 knockout mice. Expression of a mutant SNARK in mouse tibialis anterior muscle impaired contraction-stimulated, but not insulin-stimulated, glucose transport. Whole-body SNARK heterozygotic knockout mice also had impaired contraction-stimulated glucose transport in skeletal muscle, and knockdown of SNARK in C2C12 muscle cells impaired sorbitol-stimulated glucose transport. SNARK is activated by muscle contraction and is a unique mediator of contraction-stimulated glucose transport in skeletal muscle.