Human midbrain dopaminergic progenitors.

Human midbrain dopaminergic progenitors (mDAPs) are one of the most representative cell types in both basic research and clinical applications. However, there are still many challenges for the preparation and quality control of mDAPs, such as the lack of standards. Therefore, the establishment of critical quality attributes and technical specifications for mDAPs is largely needed. "Human midbrain dopaminergic progenitor" jointly drafted and agreed upon by experts from the Chinese Society for Stem Cell Research, is the first guideline for human mDAPs in China. This standard specifies the technical requirements, test methods, inspection rules, instructions for usage, labelling requirements, packaging requirements, storage requirements, transportation requirements and waste disposal requirements for human mDAPs, which is applicable to the quality control for human mDAPs. It was originally released by the China Society for Cell Biology on 30 August 2022. We hope that the publication of this guideline will facilitate the institutional establishment, acceptance and execution of proper protocols, and accelerate the international standardization of human mDAPs for clinical development and therapeutic applications.

Requirements for human natural killer cells.

'Requirements for Human Natural Killer Cells' is the latest set of guidelines on human NK cells in China, jointly drafted and agreed upon by experts from the Standards Committee of Chinese Society for Cell Biology. This standard specifies requirements for the human natural killer (NK) cells, including the technical requirements, test methods, test regulations, instructions for use, labeling requirements, packaging requirements, storage and transportation requirements, and waste disposal requirements of NK cells. This standard is applicable for the quality control of NK cells, derived from human tissues, or differentiated/transdifferentiated from stem cells. It was originally released by the Chinese Society for Cell Biology on 30 August, 2022. We hope that the publication of these guidelines will promote institutional establishment, acceptance, and execution of proper protocols and accelerate the international standardization of human NK cells for applications.

Gestational cadmium exposure disrupts fetal liver development via repressing estrogen biosynthesis in placental trophoblasts.

Cadmium (Cd), commonly found in diet and drinking water, is known to be harmful to the human liver. Nevertheless, the effects and mechanisms of gestational Cd exposure on fetal liver development remain unclear. Here, we reported that gestational Cd (150 mg/L) exposure obviously downregulated the expression of critical proteins including PCNA, Ki67 and VEGF-A in proliferation and angiogenesis in fetal livers, and lowered the estradiol concentration in fetal livers and placentae. Maternal estradiol supplement alleviated aforesaid impairments in fetal livers. Our data showed that the levels of pivotal estrogen synthases, such as CYP17A1 and 17ß-HSD, was markedly decreased in Cd-stimulated placentae but not fetal livers. Ground on ovariectomy (OVX), we found that maternal ovarian-derived estradiol had no major effects on Cd-impaired development in fetal liver. In addition, Cd exposure activated placental PERK signaling, and inhibited PERK activity could up-regulated the expressions of CYP17A1 and 17ß-HSD in placental trophoblasts. Collectively, gestational Cd exposure inhibited placenta-derived estrogen synthesis via activating PERK signaling, and therefore impaired fetal liver development. This study suggests a protective role for placenta-derived estradiol in fetal liver dysplasia shaped by toxicants, and provides a theoretical basis for toxicants to impede fetal liver development by disrupting the placenta-fetal-liver axis.

Requirements for human-induced pluripotent stem cells.

'Requirements for Human-Induced Pluripotent Stem Cells' is the first set of guidelines on human-induced pluripotent stem cells in China, jointly drafted and agreed upon by experts from the Chinese Society for Stem Cell Research. This standard specifies the technical requirements, test methods, and instructions for use, labeling, packaging, storage, transportation, and waste handling for human-induced pluripotent stem cells, which apply to the production and quality control of human-induced pluripotent stem cells. It was released by the Chinese Society for Cell Biology on 9 January 2021 and came into effect on 9 April 2021. We hope that the publication of these guidelines will promote institutional establishment, acceptance, and execution of proper protocols and accelerate the international standardization of human-induced pluripotent stem cells for applications.

Human retinal pigment epithelial cells.

'Human retinal pigment epithelial cells' is the first set of guidelines on human retinal pigment epithelial cells in China, jointly drafted and agreed upon by experts from the Chinese Society for Stem Cell Research. This standard specifies technical requirements, test methods, inspection rules, instructions for usage, labelling requirements, packaging requirements, storage requirements and transportation requirements and waste disposal requirements for human retinal pigment epithelial cells, which is applicable to quality control during the process of manufacturing and testing of human retinal pigment epithelial cells. It was originally released by the Chinese Society for Cell Biology on 9 January 2021. We hope that publication of these guidelines will promote institutional establishment, acceptance and execution of proper protocols and accelerate the international standardization of human retinal pigment epithelial cells for applications.

Requirments for primary human hepatocyte.

'Requirements for Primary Human Hepatocyte' is the first set of guidelines on Primary Human Hepatocyte in China, jointly drafted and agreed upon by experts from the Chinese Society for Stem Cell Research. This standard specifies the technical requirements, test methods, test regulations, instructions for use, labelling requirements, packaging requirements, storage requirements and transportation requirements for Primary Human Hepatocyte, which is applicable to the quality control for Primary Human Hepatocyte. It was originally released by the China Society for Cell Biology on 9 January 2021. We hope that publication of these guidelines will promote institutional establishment, acceptance and execution of proper protocols and accelerate the international standardization of Primary Human Hepatocyte for applications.

Requirements for human haematopoietic stem/progenitor cells.

'Requirements for human haematopoietic stem/progenitor cells' is the first set of guidelines on human haematopoietic stem/progenitor cells in China, jointly drafted and agreed upon by experts from the Chinese Society for Stem Cell Research. This standard specifies the technical requirements, inspection methods, inspection rules, instructions for usage, labelling requirements, packaging requirements, storage requirements and transportation requirements for human haematopoietic stem/progenitor cells, which is applicable to the quality control for human haematopoietic stem/progenitor cells. We hope that publication of these guidelines will promote institutional establishment, acceptance and execution of proper protocols, and accelerate the international standardization of human haematopoietic stem/progenitor cells for applications.

Human mesenchymal stem cells.

Mesenchymal stem cells (MSCs) have attracted great interest for cell therapy and tissue regeneration due to their self-renewal capacity, multipotency and potent immunomodulatory effects on immune cells. However, heterogeneity of MSCs has become a prominent obstacle to limit their translation into practice, as cells from different tissue sources or each individual have great differences in their transcriptomic signatures, differentiation potential and biological functions. Therefore, there is an urgent need for consensus standard for the quality control and technical specifications of MSCs. 'Human Mesenchymal Stem Cells' is the latest set of guidelines on hMSC in China, jointly drafted and agreed upon by experts from the Chinese Society for Stem Cell Research. This standard specifies the technical requirements, test methods, test regulations, instructions for use, labelling requirements, packaging requirements, storage requirements, transportation requirements and waste disposal requirements for hMSC, which is applicable to the quality control for hMSC. It was originally released by the China Society for Cell Biology on 9 January 2021. We hope that publication of these guidelines will facilitate institutional establishment, acceptance and execution of proper protocols, and accelerate the international standardization of hMSC for clinical development and therapeutic applications.

Requirements for human cardiomyocytes.

'Requirements for human cardiomyocytes', jointly drafted and agreed upon by experts from the Chinese Society for Stem Cell Research, is the first guideline for human cardiomyocytes in China. This standard specifies the technical requirements, test methods, test regulations, instructions for use, labelling requirements, packing requirements, storage requirements, transportation requirements and waste disposal requirements for human cardiomyocytes, which is designed to normalize and standardize human cardiomyocyte research and production. It was originally released by the China Society for Cell Biology on 9 January 2021. We hope that the publication of this guideline will promote institutional establishment, acceptance and execution of proper protocols, and accelerate the international standardization of human cardiomyocytes for applications.

Requirements for human embryonic stem cells.

'Requirements for Human Embryonic Stem Cells' is the first set of guidelines on human embryonic stem cells in China, jointly drafted and agreed upon by experts from the Chinese Society for Stem Cell Research. This standard specifies the technical requirements, test methods, test regulations, instructions for use, labelling requirements, packaging requirements, storage requirements and transportation requirements for human embryonic stem cells, which is applicable to the quality control for human embryonic stem cells. It was originally released by the China Society for Cell Biology on 26 February 2019 and was further revised on 30 April 2020. We hope that publication of these guidelines will promote institutional establishment, acceptance and execution of proper protocols, and accelerate the international standardization of human embryonic stem cells for applications.

Facile synthesis of a BCN nanofiber and its ultrafast adsorption performance.

Boron carbonitride (BCN) nanofibers with rapid and efficient adsorption performance were prepared by electrospinning technology. TEM, XRD, XPS and N2 adsorption-desorption isotherms were performed to study the microstructure of the nanofibers. The results showed that the BCN fibers synthesized at 1000 °C (BCN-1000) have good crystallinity and high specific surface areas (403 m2 g-1). BCN-1000 nanofibers adsorb 70% of amino black 10B (AB-10B) within 10 minutes and reach adsorption equilibrium within 60 minutes. Compared with previous reports, it is found that the adsorption rate of BCN-1000 nanofibers to amino black (AB-10B) is much higher than that of other adsorbents. And BCN nanofibers exhibit a large adsorption capacity (625 mg g-1). In addition, the process of AB-10B adsorption on BCN nanofibers was systematically investigated, which was in accordance with the pseudo-second-order kinetics model and Langmuir isotherm model.

[Heavy Metal Content of Rural Living Solid Waste and Related Source and Distribution Analysis].

Living solid waste of 72 typical villages and towns in 12 provinces was investigated, and related heavy metal pollution characteristics, source, and distribution were analyzed. Results showed that heavy metal content of As, Hg, Pb, Cd, Cr, Cu, Zn, and Ni in living solid waste of typical northern villages of China was (7.51±8.89), (0.64±0.42), (21.91±12.29), (4.82±8.37), (86.36±59.99), (36.43±15.98), (62.19±36.61), and (46.07±25.22) mg·kg-1, respectively. Content of As, Hg, Pb, Cd, Cr, Cu, Zn, and Ni in living solid waste of typical southern towns was (7.43±8.82), (0.83±0.74), (21.62±13.76), (1.84±4.55), (131.06±74.96), (37.20±16.80), (98.04±63.71), and (46.75±25.75) mg·kg-1, respectively. Cd and Hg exceeded the standards for urban garbage agricultural control and soil environmental quality. Sources of heavy metals in domestic waste were explored by Pearson correlation analysis, cluster analysis, and principal component analysis. Results showed that Pb and Cd were mainly derived from kitchen waste, dust, paper, rubber, and plastic. Hg was mainly from kitchen waste and dust. Zn and Cr were mainly from dust. Cu was mainly from dust, paper, rubber, plastic, battery, and electronic waste. Ni was mainly from battery and electronic wastes. As was mainly derived from pesticides and fertilizers.

Investigation of the physical and chemical characteristics of rural solid waste in China and its spatiotemporal distributions.

Despite governmental efforts toward the development of policies, funds, and technologies, the inherent characteristics of rural solid waste (RSW) discharge have led to great difficulties in RSW pollution control. However, establishing a realistic management strategy requires greater knowledge of RSW generation. Therefore, the RSW of 72 typical towns and villages from 12 provinces of China was analyzed for physicochemical characteristics, as well as its spatiotemporal distribution. The largest proportion of kitchen waste, coal ash, plastic, and paper of RSW was 33.70% ± 17.87%, 26.50% ± 17.61%, 13.48% ± 5.68%, and 10.75% ± 5.75%, respectively, in 2015. Although RSW had the potential for composting, it was still necessary to pay special attention to heavy metals pollution of RSW. The spatiotemporal distributions of RSW components were extremely non-homogenous, and significant variations existed in the kitchen residue, coal ash, plastic, and paper because of differences in economic growth, climatic changes, dietary habits, energy consumption structure, and consumer preferences. No obvious differences in RSW components were observed between villages and market towns. Overall, RSW treatment and management approaches should be considered based on local conditions of RSW generation.

Modification of chitin with high adsorption capacity for methylene blue removal.

Porous chitin sorbents (PChs) with different content of chitin, ranging from 0.9% to 3.5%, were prepared by gel method with CaBr2·xH2O/CH3OH solution and characterized by FT-IR, XRD and SEM. The adsorption isotherms and kinetic analysis of methylene blue (MB) onto PChs were studied. Experimental results illustrated lower crystallinity and more pores of PChs containing 3.5% chitin displayed higher adsorption capacity, the removal of MB was 79.8%. The adsorption equilibrium isotherm curve of MB onto PChs adsorbents conformed to the Freundlich equation. The PFO, PSO and Weber-Morris models were applied to fit with the adsorption kinetics. The results demonstrated the adsorption of MB might be the mass transfer of heterogeneous system and involve multiple diffusion steps. The adsorption capacity of PChs with 3.5% chitin can maintain 65% removal ratio of MB after being used six adsorption-desorption cycles. It was supposed that PChs may be a promising, cheap, environmentally friendly and efficient adsorbent for some dye wastewater treatment in the near future.

Pompe disease results in a Golgi-based glycosylation deficit in human induced pluripotent stem cell-derived cardiomyocytes.

Infantile-onset Pompe disease is an autosomal recessive disorder caused by the complete loss of lysosomal glycogen-hydrolyzing enzyme acid α-glucosidase (GAA) activity, which results in lysosomal glycogen accumulation and prominent cardiac and skeletal muscle pathology. The mechanism by which loss of GAA activity causes cardiomyopathy is poorly understood. We reprogrammed fibroblasts from patients with infantile-onset Pompe disease to generate induced pluripotent stem (iPS) cells that were differentiated to cardiomyocytes (iPSC-CM). Pompe iPSC-CMs had undetectable GAA activity and pathognomonic glycogen-filled lysosomes. Nonetheless, Pompe and control iPSC-CMs exhibited comparable contractile properties in engineered cardiac tissue. Impaired autophagy has been implicated in Pompe skeletal muscle; however, control and Pompe iPSC-CMs had comparable clearance rates of LC3-II-detected autophagosomes. Unexpectedly, the lysosome-associated membrane proteins, LAMP1 and LAMP2, from Pompe iPSC-CMs demonstrated higher electrophoretic mobility compared with control iPSC-CMs. Brefeldin A induced disruption of the Golgi in control iPSC-CMs reproduced the higher mobility forms of the LAMPs, suggesting that Pompe iPSC-CMs produce LAMPs lacking appropriate glycosylation. Isoelectric focusing studies revealed that LAMP2 has a more alkaline pI in Pompe compared with control iPSC-CMs due largely to hyposialylation. MALDI-TOF-MS analysis of N-linked glycans demonstrated reduced diversity of multiantennary structures and the major presence of a trimannose complex glycan precursor in Pompe iPSC-CMs. These data suggest that Pompe cardiomyopathy has a glycan processing abnormality and thus shares features with hypertrophic cardiomyopathies observed in the congenital disorders of glycosylation.

Observation on the ion association equilibria in NaNO3 droplets using micro-Raman spectroscopy.

Ion association ratios as a function of concentration were estimated in single NaNO(3) droplets (5-60 µm) on a polytetrafluoroethylene (PTFE) substrate with molar water-to-solute ratios (WSRs) of 0.8-28 and bulk NaNO(3) solutions with WSRs of 35-200 by combining micro-Raman spectroscopy and component band analysis. Concentrations of the NaNO(3) droplets were accurately controlled by adjusting relative humidity (RH) in a sample chamber. As the WSRs decreased from 200 to 0.8, symmetric stretching band (ν(1)-NO(3)(-)) was observed to shift from 1047 to 1058 cm(-1) along with a change in full width at half-maximum (fwhm) from ∼10 to ∼16 cm(-1), indicative of formation of ion pairs with different structures. Through the component band analysis of the ν(1)-NO(3)(-) band, five bands centered at 1040.0, 1042.9, 1048.5, 1053.5, and 1057.0 cm(-1) were identified and assigned to coupled wagging modes of water molecules hydrated with nitrate ions, free hydrated nitrate anions, solvent-shared ion pairs (SIPs), contact ion pairs (CIPs), and the complex ion aggregates (CIAs), respectively. There were large amounts of SIPs and CIPs in dilute NaNO(3) solution even at an extremely low concentration (WSR ∼ 200), and each accounted for 50% and 20% of total nitrate species, respectively. This finding is in good agreement with earlier reported observations. In the dilute solutions (45 < WSR < 200), there is the same amount of free hydrated ions transformed into SIPs as that of SIPs transformed into CIPs. As a result, the overall amount of SIPs remained unchanged over the concentration range. With a decrease in WSR from 45 to 0.8, the amounts of SIPs and free solvated NO(3)(-) ions kept decreasing, whereas the amount of CIPs rose to a maximum at WSR = 7 and then fell with a further decreasing WSR. Formation of CIAs started at WSR ∼ 45, and its amount continuously increased as the WSR is further reduced to 0.8. The effect of temperature on ion association structure in the NaNO(3) droplets was also studied in the present work. An increase in temperature promoted formation of both CIPs and CIAs, and the latter was more pronounced. At 80 °C, the most concentrated NaNO(3) droplets had a WSR approximately equal to 0.12 and were in amorphous state with cations and anions aggregated in a complicated manner, highly similar to ionic liquid.

Extracellular matrix promotes highly efficient cardiac differentiation of human pluripotent stem cells: the matrix sandwich method.

RATIONALE: Cardiomyocytes (CMs) differentiated from human pluripotent stem cells (PSCs) are increasingly being used for cardiovascular research, including disease modeling, and hold promise for clinical applications. Current cardiac differentiation protocols exhibit variable success across different PSC lines and are primarily based on the application of growth factors. However, extracellular matrix is also fundamentally involved in cardiac development from the earliest morphogenetic events, such as gastrulation. OBJECTIVE: We sought to develop a more effective protocol for cardiac differentiation of human PSCs by using extracellular matrix in combination with growth factors known to promote cardiogenesis. METHODS AND RESULTS: PSCs were cultured as monolayers on Matrigel, an extracellular matrix preparation, and subsequently overlayed with Matrigel. The matrix sandwich promoted an epithelial-to-mesenchymal transition as in gastrulation with the generation of N-cadherin-positive mesenchymal cells. Combining the matrix sandwich with sequential application of growth factors (Activin A, bone morphogenetic protein 4, and basic fibroblast growth factor) generated CMs with high purity (up to 98%) and yield (up to 11 CMs/input PSC) from multiple PSC lines. The resulting CMs progressively matured over 30 days in culture based on myofilament expression pattern and mitotic activity. Action potentials typical of embryonic nodal, atrial, and ventricular CMs were observed, and monolayers of electrically coupled CMs modeled cardiac tissue and basic arrhythmia mechanisms. CONCLUSIONS: Dynamic extracellular matrix application promoted epithelial-mesenchymal transition of human PSCs and complemented growth factor signaling to enable robust cardiac differentiation.

Suppression of NaNO3 crystal nucleation by glycerol: micro-Raman observation on the efflorescence process of mixed glycerol/NaNO3/water droplets.

Although the hygroscopicity of a NaNO(3)/water microdroplet and a polyalcohol/water microdroplet, two of the most important aerosols in atmosphere, has been widely studied, little is known about the relationship between the hygroscopic behavior of mixed NaNO(3)/polyalcohol/water droplets and their structures on the molecular level. In this study, the hygroscopicity of mixed glycerol/NaNO(3)/water droplets deposited on a hydrophobic substrate was studied by micro-Raman spectroscopy with organic-to-inorganic molar ratios (OIRs) of 0.5, 1, and 2. In the mixed glycerol/NaNO(3)/water droplets, glycerol molecules tended to combine with Na(+) and NO(3)(-) ions by electrostatic interaction and hydrogen bonding, respectively. On the basis of the analyses of the changes of symmetric stretching (v(s)-CH(2)), asymmetric stretching (v(a)-CH(2)), their area ratio (Av(a)-CH(2)/Av(s)-CH(2)) of glycerol, and symmetric stretching band of NO(3)(-) (ν(1)-NO(3)(-)) with relative humidity (RH), it was found that the conformation of glycerol was transformed from αα mainly to γγ and partly to αγ with a decreasing RH in the mixed droplets, contrary to the case in the glycerol/water droplet. In addition, the glycerol with γγ and αγ conformation had strong interaction with Na(+) and NO(3)(-) respectively, which suppressed the formation of contact of ions and delayed the efflorescence relative humidity (ERH) for the mixed droplets compared to the NaNO(3)/water droplet.

Protein kinase C mediated extraembryonic endoderm differentiation of human embryonic stem cells.

Unlike mouse embryonic stem cells (ESCs), which are closely related to the inner cell mass, human ESCs appear to be more closely related to the later primitive ectoderm. For example, human ESCs and primitive ectoderm share a common epithelial morphology, growth factor requirements, and the potential to differentiate to all three embryonic germ layers. However, it has previously been shown that human ESCs can also differentiate to cells expressing markers of trophoblast, an extraembryonic lineage formed before the formation of primitive ectoderm. Here, we show that phorbol ester 12-O-tetradecanoylphorbol 13-acetate causes human ESCs to undergo an epithelial mesenchymal transition and to differentiate into cells expressing markers of parietal endoderm, another extraembryonic lineage. We further confirmed that this differentiation is through the activation of protein kinase C (PKC) pathway and demonstrated that a particular PKC subtype, PKC-δ, is most responsible for this transition.

Human lymphoblastoid B-cell lines reprogrammed to EBV-free induced pluripotent stem cells.

Generation of patient-specific induced pluripotent cells (iPSCs) holds great promise for regenerative medicine. Epstein-Barr virus immortalized lymphoblastoid B-cell lines (LCLs) can be generated from a minimal amount of blood and are banked worldwide as cellular reference material for immunologic or genetic analysis of pedigreed study populations. We report the generation of iPSCs from 2 LCLs (LCL-iPSCs) via a feeder-free episomal method using a cocktail of transcription factors and small molecules. LCL-derived iPSCs exhibited normal karyotype, expressed pluripotency markers, lost oriP/EBNA-1 episomal vectors, generated teratomas, retained donor identity, and differentiated in vitro into hematopoietic, cardiac, neural, and hepatocyte-like lineages. Significantly, although the parental LCLs express viral EBNA-1 and other Epstein-Barr virus latency-related elements for their survival, their presence was not detectable in LCL-iPSCs. Thus, reprogramming LCLs could offer an unlimited source for patient-specific iPSCs.