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.

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.

Elucidation of HHEX in pancreatic endoderm differentiation using a human iPSC differentiation model.

For pluripotent stem cell (PSC)-based regenerative therapy against diabetes, the differentiation efficiency to pancreatic lineage cells needs to be improved based on the mechanistic understanding of pancreatic differentiation. Here, we aimed to elucidate the molecular mechanisms underlying pancreatic endoderm differentiation by searching for factors that regulate a crucial pancreatic endoderm marker gene, NKX6.1. Unbiasedly screening an siRNA knockdown library, we identified a candidate transcription factor, HHEX. HHEX knockdown suppressed the expression of another pancreatic endoderm marker gene, PTF1A, as well as NKX6.1, independently of PDX1, a known regulator of NKX6.1 expression. In contrast, the overexpression of HHEX upregulated the expressions of NKX6.1 and PTF1A. RNA-seq analysis showed decreased expressions of several genes related to pancreatic development, such as NKX6.1, PTF1A, ONECUT1 and ONECUT3, in HHEX knockdown pancreatic endoderm. These results suggest that HHEX plays a key role in pancreatic endoderm differentiation.

CDK8/19 inhibition plays an important role in pancreatic ß-cell induction from human iPSCs.

BACKGROUND: Transplantation of differentiated cells from human-induced pluripotent stem cells (hiPSCs) holds great promise for clinical treatments. Eliminating the risk factor of malignant cell transformation is essential for ensuring the safety of such cells. This study was aimed at assessing and mitigating mutagenicity that may arise during the cell culture process in the protocol of pancreatic islet cell (iPIC) differentiation from hiPSCs. METHODS: We evaluated the mutagenicity of differentiation factors used for hiPSC-derived pancreatic islet-like cells (iPICs). We employed Ames mutagenicity assay, flow cytometry analysis, immunostaining, time-resolved fluorescence resonance energy transfer-based (TR-FRET) cell-free dose-response assays, single-cell RNA-sequencing and in vivo efficacy study. RESULTS: We observed a mutagenic effect of activin receptor-like kinase 5 inhibitor II (ALK5iII). ALK5iII is a widely used ß-cell inducer but no other tested ALK5 inhibitors induced ß-cells. We obtained kinase inhibition profiles and found that only ALK5iII inhibited cyclin-dependent kinases 8 and 19 (CDK8/19) among all ALK5 inhibitors tested. Consistently, CDK8/19 inhibitors efficiently induced ß-cells in the absence of ALK5iII. A combination treatment with non-mutagenic ALK5 inhibitor SB431542 and CDK8/19 inhibitor senexin B afforded generation of iPICs with in vitro cellular composition and in vivo efficacy comparable to those observed with ALK5iII. CONCLUSION: Our findings suggest a new risk mitigation approach for cell therapy and advance our understanding of the ß-cell differentiation mechanism.

Purification of human iPSC-derived cells at large scale using microRNA switch and magnetic-activated cell sorting.

For regenerative cell therapies using pluripotent stem cell (PSC)-derived cells, large quantities of purified cells are required. Magnetic-activated cell sorting (MACS) is a powerful approach to collect target antigen-positive cells; however, it remains a challenge to purify various cell types efficiently at large scale without using antibodies specific to the desired cell type. Here we develop a technology that combines microRNA (miRNA)-responsive mRNA switch (miR-switch) with MACS (miR-switch-MACS) to purify large amounts of PSC-derived cells rapidly and effectively. We designed miR-switches that detect specific miRNAs expressed in target cells and controlled the translation of a CD4-coding transgene as a selection marker for MACS. For the large-scale purification of induced PSC-derived cardiomyocytes (iPSC-CMs), we transferred miR-208a-CD4 switch-MACS and obtained purified iPSC-CMs efficiently. Moreover, miR-375-CD4 switch-MACS highly purified pancreatic insulin-producing cells and their progenitors expressing Chromogranin A. Overall, the miR-switch-MACS method can efficiently purify target PSC-derived cells for cell replacement therapy.

Biofeedback Physical Therapy With the Hybrid Assistive Limb (HAL) Lumbar Type for Chronic Low Back Pain: A Pilot Study.

Objective There are many treatments for chronic low back pain, including various medications, exercise therapy, orthotics, and surgery, but no treatment is definitive. We hypothesized that biofeedback therapy using the hybrid assistive limb (HAL) lumbar type would have some immediate effects on chronic low back pain. The purpose of this pilot study was to assess whether immediate changes in low back pain and hip flexibility and any other adverse events would occur following the HAL biofeedback physical therapy. Methods This was a single-center, pilot, prospective, single-arm study of outpatient biofeedback physical therapy using the HAL lumbar type for patients with chronic low back pain. Patients underwent a 10-minute biofeedback physical therapy (lumbar flexion-extension, sit-to-stand, and squat) with the HAL lumbar type (in one session). The visual analog scale (VAS) score of low back pain during lumbar flexion, extension, lateral bending, and rotation was evaluated. The finger-to-floor distance (FFD), straight leg raising test (SLR), and the Thomas test were measured to assess hip flexibility. Results All 35 participants (14 men and 21 women) (100%) conducted a biofeedback HAL therapy session using the HAL lumbar type. No participant had deterioration of low back pain. No adverse events occurred. After the biofeedback therapy using the HAL lumbar type, SLR demonstrated a significant positive change with large effect size and sufficient power. Lumbar VAS during lumbar flexion and extension and FFD showed a significant positive change with medium effect size and adequate power. Conclusions Biofeedback therapy using the HAL lumbar type is an option for intervention in chronic low back pain.

Microwell bag culture for large-scale production of homogeneous islet-like clusters.

Pluripotent stem-cell derived cells can be used for type I diabetes treatment, but we require at least 105-106 islet-like clusters per patient. Although thousands of uniform cell clusters can be produced using a conventional microwell plate, numerous obstacles need to be overcome for its clinical use. In this study, we aimed to develop a novel bag culture method for the production of uniform cell clusters on a large scale (105-106 clusters). We prepared small-scale culture bags (< 105 clusters) with microwells at the bottom and optimized the conditions for producing uniform-sized clusters in the bag using undifferentiated induced pluripotent stem cells (iPSCs). Subsequently, we verified the suitability of the bag culture method using iPSC-derived pancreatic islet cells (iPICs) and successfully demonstrate the production of 6.5 × 105 uniform iPIC clusters using a large-scale bag. In addition, we simplified the pre- and post-process of the culture-a degassing process before cell seeding and a cluster harvesting process. In conclusion, compared with conventional methods, the cluster production method using bags exhibits improved scalability, sterility, and operability for both clinical and research use.

Characterization and reduction of non-endocrine cells accompanying islet-like endocrine cells differentiated from human iPSC.

The differentiation of pancreatic endocrine cells from human pluripotent stem cells has been thoroughly investigated for their application in cell therapy against diabetes. Although non-endocrine cells are inevitable contaminating by-products of the differentiation process, a comprehensive profile of such cells is lacking. Therefore, we characterized non-endocrine cells in iPSC-derived pancreatic islet cells (iPIC) using single-cell transcriptomic analysis. We found that non-endocrine cells consist of (1) heterogeneous proliferating cells, and (2) cells with not only pancreatic traits but also liver or intestinal traits marked by FGB or AGR2. Non-endocrine cells specifically expressed FGFR2, PLK1, and LDHB. We demonstrated that inhibition of pathways involving these genes selectively reduced the number of non-endocrine cells in the differentiation process. These findings provide useful insights into cell purification approaches and contribute to the improvement of the mass production of endocrine cells for stem cell-derived cell therapy for diabetes.

Combined Omics Approaches Reveal the Roles of Non-canonical WNT7B Signaling and YY1 in the Proliferation of Human Pancreatic Progenitor Cells.

The proliferation of human pancreatic progenitor cells (PPCs) is critical for developing cell therapies for diabetes. Here, using transcriptome analysis combined with small interfering RNA (siRNA) screening, we revealed that WNT7B is a downstream growth factor of AT7867, a compound known to promote the proliferation of PPCs generated from human pluripotent stem cells. Feeder cell lines stably expressing mouse Wnt7a or Wnt7b, but not other Wnts, enhanced PPC proliferation in the absence of AT7867. Importantly, Wnt7a/b ligands did not activate the canonical Wnt pathway, and PPC proliferation depended on the non-canonical Wnt/PKC pathway. A comparison of the phosphoproteome in response to AT7867 or a newly synthesized AT7867 derivative uncovered the function of YY1 as a transcriptional regulator of WNT7B. Overall, our data highlight unknown roles of non-canonical WNT7B/PKC signaling and YY1 in human PPC proliferation and will contribute to the stable supply of a cell source for pancreatic disease modeling and therapeutic applications.

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.

Enhanced Device Performance with Passivation of the TiO2 Surface Using a Carboxylic Acid Fullerene Monolayer for a SnPb Perovskite Solar Cell with a Normal Planar Structure.

Research on tin-lead (SnPb) perovskite solar cells (PSCs) has gained popularity in recent years because of their low band gap, which could be applied to tandem solar cells. However, most of the work is based on inverted PSCs using PEDOT:PSS as the hole-transport layer as normal-structure PSCs show lower efficiency. In this work, the reason behind the low efficiency of normal-structure SnPb PSCs is elucidated and surface passivation has been tested as a method to overcome the problem. In the case of normal PSCs, at the interface between the titania layer and SnPb perovskite, there are many carrier traps observed originating from Ti-O-Sn bonds. In order to avoid the direct contact between titania and the SnPb perovskite layer, the titania surface is passivated with carboxylic acid C60 resulting in an efficiency increase from 5.14 to 7.91%. This will provide a direction of enhancing the efficiency of the normal-structure SnPb PSCs through heterojunction engineering.

Insulin-Deficient Diabetic Condition Upregulates the Insulin-Secreting Capacity of Human Induced Pluripotent Stem Cell-Derived Pancreatic Endocrine Progenitor Cells After Implantation in Mice.

The host environment is a crucial factor for considering the transplant of stem cell-derived immature pancreatic cells in patients with type 1 diabetes. Here, we investigated the effect of insulin (INS)-deficient diabetes on the fate of immature pancreatic endocrine cell grafts and the underlying mechanisms. Human induced pluripotent stem cell-derived pancreatic endocrine progenitor cells (EPCs), which contained a high proportion of chromogranin A+ NK6 homeobox 1+ cells and very few INS+ cells, were used. When the EPCs were implanted under the kidney capsule in immunodeficient mice, INS-deficient diabetes accelerated increase in plasma human C-peptide, a marker of graft-derived INS secretion. The acceleration was suppressed by INS infusion but not affected by partial attenuation of hyperglycemia by dapagliflozin, an INS-independent glucose-lowering agent. Immunohistochemical analyses indicated that the grafts from diabetic mice contained more endocrine cells including proliferative INS-producing cells compared with that from nondiabetic mice, despite no difference in whole graft mass between the two groups. These data suggest that INS-deficient diabetes upregulates the INS-secreting capacity of EPC grafts by increasing the number of endocrine cells including INS-producing cells without changing the graft mass. These findings provide useful insights into postoperative diabetic care for cell therapy using stem cell-derived pancreatic cells.

Relationship between Lattice Strain and Efficiency for Sn-Perovskite Solar Cells.

In the composition of Q0.1(FA0.75MA0.25)0.9SnI3, Q is replaced with Na+, K+, Cs+, ethylammonium+ (EA+), and butylammonium+ (BA+), respectively, and the relationship between actually measured lattice strain and photovoltaic performances is discussed. The lattice strain evaluated by the Williamson-hall plot of X-ray diffraction data decreased as the tolerance factor was close to one. The efficiency of the Sn-perovskite solar cell was enhanced as the lattice strain decreased. Among them, EA0.1(FA0.75MA0.25)0.9SnI3 having lowest lattice strain gave the best result of 5.41%. Because the carrier mobility increased with a decrease in the lattice strain, these lattice strains would disturb carrier mobility and decrease the solar cell efficiency. Finally, the results that the efficiency of the SnGe-perovskite solar cells was gradually enhanced from 6.42 to 7.60% during storage, was explained by the lattice strain relaxation during the storage.

Suppression of Charge Carrier Recombination in Lead-Free Tin Halide Perovskite via Lewis Base Post-treatment.

Lead-free tin perovskite solar cells (PSCs) show the most promise to replace the more toxic lead-based perovskite solar cells. However, the efficiency is significantly less than that of lead-based PSCs as a result of low open-circuit voltage. This is due to the tendency of Sn2+ to oxidize into Sn4+ in the presence of air together with the formation of defects and traps caused by the fast crystallization of tin perovskite materials. Here, post-treatment of the tin perovskite layer with edamine Lewis base to suppress the recombination reaction in tin halide PSCs results in efficiencies higher than 10%, which is the highest reported efficiency to date for pure tin halide PSCs. The X-ray photoelectron spectroscopy data suggest that the recombination reaction originates from the nonstoichiometric Sn:I ratio rather than the Sn4+:Sn2+ ratio. The amine group in edamine bonded the undercoordinated tin, passivating the dangling bonds and defects, resulting in suppressed charge carrier recombination.

Efficient Generation of Pancreas/Duodenum Homeobox Protein 1+ Posterior Foregut/Pancreatic Progenitors from hPSCs in Adhesion Cultures.

Human pluripotent stem cell (hPSC)-derived pancreatic cells are a promising cell source for regenerative medicine and a platform to study human developmental processes. Stepwise directed differentiation that recapitulates developmental processes is one of the major ways to generate pancreatic cells including pancreas/duodenum homeobox protein 1+ (PDX1+) pancreatic progenitor cells. Conventional protocols initiate the differentiation with small colonies shortly after the passage. However, in the state of colonies or aggregates, cells are prone to heterogeneities, which might hamper the differentiation to PDX1+ cells. Here, we present a detailed protocol to differentiate hPSCs into PDX1+ cells. The protocol consists of four steps and initiates the differentiation by seeding dissociated single cells. The induction of SOX17+ definitive endoderm cells was followed by the expression of two primitive gut tube markers, HNF1ß and HNF4α, and eventual differentiation into PDX1+ cells. The present protocol provides easy handling and may improve and stabilize the differentiation efficiency of some hPSC lines that were previously found to differentiate inefficiently into endodermal lineages or PDX1+ cells.

Differentiation and isolation of iPSC-derived remodeling ductal plate-like cells by use of an AQP1-GFP reporter human iPSC line.

Cholangiocytes are the epithelial cells that line bile ducts, and ductal plate malformation is a developmental anomaly of bile ducts that causes severe congenital biliary disorders. However, because of a lack of specific marker genes, methods for the stepwise differentiation and isolation of human induced pluripotent stem cell (hiPSC)-derived cholangiocyte progenitors at ductal plate stages have not been established. We herein generated an AQP1-GFP reporter hiPSC line and developed a combination treatment with transforming growth factor (TGF) ß2 and epidermal growth factor (EGF) to induce hiPSC-derived hepatoblasts into AQP1+ cells in vitro. By confirming that the isolated AQP1+ cells showed similar gene expression patterns to cholangiocyte progenitors at the remodeling ductal plate stage around gestational week (GW) 20, we established a differentiation protocol from hiPSCs through SOX9+CK19+AQP1- ductal plate-like cells into SOX9+CK19+AQP1+ remodeling ductal plate-like cells. We further generated 3D bile duct-like structures from the induced ductal plate-like cells. These results suggest that AQP1 is a useful marker for the generation of remodeling ductal plate cells from hiPSCs. Our methods may contribute to elucidating the differentiation mechanisms of ductal plate cells and the pathogenesis of ductal plate malformation.

The interparticle distance limit for multiple exciton dissociation in PbS quantum dot solid films.

Understanding the behaviour of multiple exciton dissociation in quantum dot (QD) solid films is of fundamental interest and paramount importance for improving the performance of quantum dot solar cells (QDSCs). Unfortunately, the charge transfer behaviour of photogenerated multiple exciton in QD solid films is not clear to date. Herein, we systematically investigate the multiple exciton charge transfer behaviour in PbS QD solid films by using ultrafast transient absorption spectroscopy. We observe that the multiple exciton charge transfer rate within QD ensembles is exponentially enhanced as the interparticle distance between the QDs decreases. Biexciton and triexciton dissociation between adjacent QDs occurs via a charge transfer tunneling effect just like single exciton, and the charge tunneling constants of the single exciton (ß1: 0.67 ± 0.02 nm-1), biexciton (ß2: 0.68 ± 0.05 nm-1) and triexciton (ß3: 0.71 ± 0.01 nm-1) are obtained. More importantly, for the first time, the interparticle distance limit (≤4.3 nm) for multiple exciton charge transfer between adjacent QDs is found for the extraction of multiple excitons rapidly before the occurrence of Auger recombination. This result points out a vital and necessary condition for the use of multiple excitons produced in PbS QD films, especially for their applications in QDSCs.

Interface Passivation Effects on the Photovoltaic Performance of Quantum Dot Sensitized Inverse Opal TiO2 Solar Cells.

Quantum dot (QD)-sensitized solar cells (QDSSCs) are expected to achieve higher energy conversion efficiency than traditional single-junction silicon solar cells due to the unique properties of QDs. An inverse opal (IO)-TiO2 (IO-TiO2) electrode is useful for QDSSCs because of its three-dimensional (3D) periodic nanostructures and better electrolyte penetration compared to the normal nanoparticles (NPs)-TiO2 (NPs-TiO2) electrode. We find that the open-circuit voltages Voc of the QDSSCs with IO-TiO2 electrodes are higher than those of QDSSCs with NPs-TiO2 electrodes. One important strategy for enhancing photovoltaic conversion efficiency of QDSSCs with IO-TiO2 electrodes is surface passivation of photoanodes using wide-bandgap semiconducting materials. In this study, we have proposed surface passivation on IO-TiO2 with ZnS coating before QD deposition. The efficiency of QDSSCs with IO-TiO2 electrodes is largely improved (from 0.74% to 1.33%) because of the enhancements of Voc (from 0.65 V to 0.74 V) and fill factor (FF) (from 0.37 to 0.63). This result indicates that ZnS passivation can reduce the interfacial recombination at the IO-TiO2/QDs and IO-TiO2/electrolyte interfaces, for which two possible explanations can be considered. One is the decrease of recombination at IO-TiO2/electrolyte interfaces, and the other one is the reduction of the back-electron injection from the TiO2 electrode to QDs. All of the above results are effective for improving the photovoltaic properties of QDSSCs.

Lead Selenide Colloidal Quantum Dot Solar Cells Achieving High Open-Circuit Voltage with One-Step Deposition Strategy.

Lead selenide (PbSe) colloidal quantum dots (CQDs) are considered to be a strong candidate for high-efficiency colloidal quantum dot solar cells (CQDSCs) due to its efficient multiple exciton generation. However, currently, even the best PbSe CQDSCs can only display open-circuit voltage ( Voc) about 0.530 V. Here, we introduce a solution-phase ligand exchange method to prepare PbI2-capped PbSe (PbSe-PbI2) CQD inks, and for the first time, the absorber layer of PbSe CQDSCs was deposited in one step by using this PbSe-PbI2 CQD inks. One-step-deposited PbSe CQDs absorber layer exhibits fast charge transfer rate, reduced energy funneling, and low trap assisted recombination. The champion large-area (active area is 0.35 cm2) PbSe CQDSCs fabricated with one-step PbSe CQDs achieve a power conversion efficiency (PCE) of 6.0% and a Voc of 0.616 V, which is the highest Voc among PbSe CQDSCs reported to date.