Human liver organoids generated with single donor-derived multiple cells rescue mice from acute liver failure

BackgroundAcute liver failure (ALF) is a life-threatening disease with a high mortality rate. However, there are limited treatments or devices available for ALF therapy. Here, we aimed to develop a new strategy for ALF treatment by transplanting functional liver organoids (LOs) generated from single donor-derived human induced pluripotent stem cell (hiPSC) endoderm, endothelial cells (ECs), and mesenchymal cells (MCs).MethodsFirst, we isolated ECs and MCs from a single donor umbilical cord (UC) through enzyme digestion and characterized the UC-ECs and UC-MCs by flow cytometry. Second, using a nonviral reprogramming method, we generated same donor-derived hiPSCs from the UC-ECs and investigated their hepatic differentiation abilities. Finally, we simultaneously plated EC-hiPSC endoderm, UC-ECs, and UC-MCs in a three-dimensional (3D) microwell culture system, and generated single donor cell-derived differentiated LOs for ALF mouse treatment.ResultsWe obtained ECs and MCs from a single donor UC with high purity, and these cells provided a multicellular microenvironment that promoted LO differentiation. hiPSCs from the same donor were generated from UC-ECs, and the resultant EC-hiPSCs could be differentiated efficiently into pure definitive endoderm and further into hepatic lineages. Simultaneous plating of EC-hiPSC endoderm, UC-ECs, and UC-MCs in the 3D microwell system generated single donor cell-derived LOs (SDC-LOs) that could be differentiated into functional LOs with enhanced hepatic capacity as compared to that of EC-hiPSC-derived hepatic-like cells. When these functional SDC-LOs were transplanted into the renal subcapsules of ALF mice, they rapidly assumed hepatic functions and improved the survival rate of ALF mice.ConclusionThese results demonstrate that functional LOs generated from single donor cells can improve the condition of ALF mice. Functional SDC-LO transplantation provides a promising novel approach for ALF therapy

Hepatic stem cells with self-renewal and liver repopulation potential are harbored in CDCP1-positive subpopulations of human fetal liver cells

BackgroundMature human hepatocytes are critical in preclinical research and therapy for liver disease, but are difficult to manipulate and expand in vitro. Hepatic stem cells (HpSCs) may be an alternative source of functional hepatocytes for cell therapy and disease modeling. Since these cells play an import role in regenerative medicine, the precise characterization that determines specific markers used to isolate these cells as well as whether they contribute to liver regeneration still remain to be shown.MethodIn this study, human HpSCs were isolated from human primary fetal liver cells (FLCs) by flow cytometry using CDCP1, CD90, and CD66 antibodies. The isolated CDCP1+CD90+CD66– HpSCs were cultured on dishes coated with type IV collagen in DMEM nutrient mixture F-12 Ham supplemented with FBS, human γ-insulin, nicotinamide, dexamethasone, and l-glutamine for at least 2 weeks, and were characterized by transcriptomic profiling, quantitative real-time PCR, immunocytochemistry, and in-vivo transplantation.ResultsThe purified CDCP1+CD90+CD66– subpopulation exhibited clonal expansion and self-renewal capability, and bipotential capacity was further identified in single cell-derived colonies containing distinct hepatocytes and cholangiocytes. Moreover, in-vivo liver repopulation assays demonstrated that human CDCP1+CD90+CD66– HpSCs repopulated over 90% of the mouse liver and differentiated into functional hepatocytes with drug metabolism activity.ConclusionsWe identified a human hepatic stem/progenitor population in the CDCP1+CD90+CD66– subpopulation in human FLCs, indicating CDCP1 marker could potentially be utilized to identify and isolate HpSCs for further cytotherapy of liver disease

(E)-N′-[1-(4-Bromo­phen­yl)ethyl­idene]benzohydrazide

The asymmetric unit of the title compound, C15H13BrN2O, contains two independent mol­ecules with different conformations; the two aromatic rings form dihedral angles of 32.4 (4) and 27.5 (4)° in the two mol­ecules. In the crystal structure, inter­molecular N—H⋯O hydrogen bonds link mol­ecules into chains propagating in [100]

Development and Comparison of Predictive Models Based on Different Types of Influencing Factors to Select the Best One for the Prediction of OSAHS Prevalence

ObjectiveThis study aims to retrospectively analyze numerous related clinical data to identify three types of potential influencing factors of obstructive sleep apnea-hypopnea syndrome (OSAHS) for establishing three predictive nomograms, respectively. The best performing one was screened to guide further clinical decision-making.MethodsCorrelation, difference and univariate logistic regression analysis were used to identify the influencing factors of OSAHS. Then these factors are divided into three different types according to the characteristics of the data. Lasso regression was used to filter out three types of factors to construct three nomograms, respectively. Compare the performance of the three nomograms evaluated by C-index, ROC curve and Decision Curve Analysis to select the best one. Two queues were obtained by randomly splitting the whole queue, and similar methods are used to verify the performance of the best nomogram.ResultsIn total, 8 influencing factors of OSAHS have been identified and divided into three types. Lasso regression finally determined 6, 3 and 4 factors to construct mixed factors nomogram (MFN), baseline factors nomogram (BAFN) and blood factors nomogram (BLFN), respectively. MFN performed best among the three and also performed well in multiple queues.ConclusionCompared with BAFN and BLFN constructed by single-type factors, MFN constructed by six mixed-type factors shows better performance in predicting the risk of OSAHS

Recapitulation of hepatitis B virus–host interactions in liver organoids from human induced pluripotent stem cells

Therapies against hepatitis B virus (HBV) have improved in recent decades; however, the development of individualized treatments has been limited by the lack of individualized infection models. In this study, we used human induced pluripotent stem cell (hiPSC) to generate a functional liver organoid (LO) that inherited the genetic background of the donor, and evaluated its application in modeling HBV infection and exploring virus–host interactions. To establish a functional hiPSC-LO, we cultured hiPSC-derived endodermal, mesenchymal, and endothelial cells with a chemically defined medium in a three-dimensional microwell culture system. Based on cell-cell interactions, these cells could organize themselves and gradually differentiate into a functional organoid, which exhibited stronger hepatic functions than hiPSC derived hepatic like cell (HLC). Moreover, the functional LO demonstrated more susceptibility to HBV infection than hiPSC-HLC, and could maintain HBV propagation and produce infectious virus for a prolonged duration. Furthermore, we found that virus infection could cause hepatic dysfunction of hiPSC-LOs, with down-regulation of hepatic gene expression, induced release of early acute liver failure markers, and altered hepatic ultrastructure. Therefore, our study demonstrated that HBV infection in hiPSC-LOs could recapitulate virus life cycle and virus induced hepatic dysfunction, suggesting that hiPSC-LOs may provide a promising individualized infection model for the development of individualized treatment for hepatitis

Assessing the concentration and potential health risk of heavy metals in China's main deciduous fruits

AbstractTo assess levels of contamination and human health risk, we analyzed the concentrations of the heavy metals lead (Pb), cadmium (Cd), chromium (Cr), and nickel (Ni) in China's main deciduous fruits — apple, pear, peach, grape, and jujube. The concentration order of the heavy metals was Ni>Cr>Pb>Cd. In 97.5% of the samples, heavy metal concentrations were within the maximum permissible limits. Among the fruits studied, the heavy metal concentrations in jujube and peach proved to be the highest, and those in grape proved to be the lowest. Only 2.2% of the samples were polluted by Ni, only 0.4% of the samples were polluted by Pb, and no samples were polluted by Cd or Cr. Compared with the other fruits, the combined heavy metal pollution was significantly higher (P<0.05) in peach and significantly lower (P<0.05) in grape. For the combined heavy metal pollution, 96.9% of the samples were at safe level, 2.32% at warning level, 0.65% at light level, and 0.13% at moderate level. In the fruits studied, the contribution of heavy metals to the daily intake rates (DIR) followed the order of Ni>Cr>Pb>Cd. The highest DIR came from apple, while the lowest DIR came from grape. For each of the heavy metals, the total DIR from five studied fruits corresponded to no more than 1.1% of the tolerable daily intake, indicating that no significant adverse health effects are expected from the heavy metals and the fruits studied. The target hazard quotients and the total target hazard quotients demonstrated that none of the analyzed heavy metals may pose risk to consumers through the fruits studied. The highest risk was posed by apple, followed in decreasing order by peach and pear, jujube, and grape. We suggest that the main deciduous fruits (apple, pear, peach, grape, and jujube) of China's main producing areas are safe to eat

Microwave-synthesized freestanding iron-carbon nanotubes on polyester composites of woven Kevlar fibre and silver nanoparticle-decorated graphene

We synthesized Ag nanoparticle-decorated multilayered graphene nanosheets (Ag-graphene) from graphite nanoplatelets and silver nitrate through 90-100&amp;#8201;s of microwave exposure, without the use of any mineral acids or harsh reducing agents. Fe nanoparticle-decorated carbon nanotubes (Fe-CNTs) were grown on polypyrrole (PPy) deposited on woven Kevlar fibre (WKF), using ferrocene as a catalyst, under microwave irradiation. Fe-CNTs grown on WKF and Ag-graphene dispersed in polyester resin (PES) were combined to fabricate Ag-graphene/Fe-CNT/PPy-coated WKF/PES composites by vacuum-assisted resin transfer moulding. The combined effect of Fe-CNTs and Ag-graphene in the resulting composites resulted in a remarkable enhancement of tensile properties (a 192.56% increase in strength and 100.64% increase in modulus) as well as impact resistance (a 116.33% increase). The electrical conductivity significantly increased for Ag-graphene/Fe-CNT/PPy-coated WKF/PES composites. The effectiveness of electromagnetic interference shielding, which relies strongly on the Ag-graphene content in the composites, was 25 times higher in Ag-graphene/Fe-CNT/PPy-coated WKF/PES than in neat WKF/PES composites. The current work offers a novel route for fabricating highly promising, cost effective WKF/PES composites through microwave-assisted synthesis of Fe-CNTs and Ag-graphene.ope