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[Display omitted] The dynamics of gas–liquid interfaces differs between aqueous surfactant and nanoparticle mixtures in rectangular cross-section capillaries. We designed and fabricated a new microfluidic device with a meter-long channel and a noncircular cross section (35 μm by 100 μm by 1 m) to study the flow behavior of long bubbles in capillaries wetted by water as well as surfactant and nanoparticle solutions. Flow in the novel symmetric loop device maintains essentially straight and inertialess conditions over a wide range of flow rates. The pressure-drop versus velocity relationship of long bubbles in capillaries with noncircular cross sections has been studied theoretically but not extensively validated. The measured pressure drop (normalized with respect to interfacial properties) experienced by bubbles varies as C a 2 / 3 over the range 10 - 7 < C a < 10 - 4 where Ca = μ U σ. Thus, the drag also scales as C a 2 / 3 . The measurements from our new device thereby confirm previous theoretical relationships of the flow of long gas bubbles wetted by surfactant solutions in noncircular capillaries. Contrary to the hypothesis, the C a 2 / 3 drag–velocity relationship applies to gas bubbles in deionized water, nanoparticles in brine, surfactant in brine, and nanoparticle-surfactant mixtures in brine for the conditions studied. [ABSTRACT FROM AUTHOR]

Background: Human keratinocytes and derived products are crucial for skin repair and regeneration. Despite substantial advances in engineered skin equivalents, their poor availability and immunorejection remain major challenges in skin grafting. Methods: Induced keratinocyte-like cells (iKCs) were directly reprogrammed from human urine cells by retroviral transduction of two lineage-specific transcription factors BMI1 and △NP63α (BN). Expression of keratinocyte stem cell or their differentiation markers were assessed by PCR, immunofluorescence and RNA-Sequencing. Regeneration capacity of iKCs were assessed by reconstitution of a human skin equivalent under air-interface condition. Results: BN-driven iKCs were similar to primary keratinocytes (pKCs) in terms of their morphology, protein expression, differentiation potential, and global gene expression. Moreover, BN-iKCs self-assembled to form stratified skin equivalents in vitro. Conclusions: This study demonstrated an approach to generate human iKCs that could be directly reprogrammed from human somatic cells and extensively expanded in serum- and feeder cell-free systems, which will facilitate their broad applicability in an efficient and patient-specific manner. [ABSTRACT FROM AUTHOR]

Engraftment of oligodendrocyte progenitor cells (OPCs), which form myelinating oligodendrocytes, has the potential to treat demyelinating diseases such as multiple sclerosis. However, conventional strategies for generating oligodendrocytes have mainly focused on direct differentiation into forebrain- or spinal cord–restricted oligodendrocytes without establishing or amplifying stem/progenitor cells. Taking advantage of a recently established culture system, we generated expandable EN1- and GBX2-positive glial-restricted progenitor-like cells (GPLCs) near the anterior hindbrain. These cells expressed PDGFRα, CD9, S100β, and SOX10 and mostly differentiated into GFAP-positive astrocytes and MBP-positive oligodendrocytes. RNA-seq analysis revealed that the transcriptome of GPLCs was similar to that of O4-positive OPCs, but distinct from that of rosette-type neural stem cells. Notably, engrafted GPLCs not only differentiated into GFAP-positive astrocytes but also myelinated the brains of adult shiverer mice 8 weeks after transplantation. Our strategy for establishing anterior hindbrain-specific GPLCs with gliogenic potency will facilitate their use in the treatment of demyelinating diseases and studies of the molecular mechanisms underlying glial development in the hindbrain. [ABSTRACT FROM AUTHOR]

Silicon-based microfluidic devices, so-called micromodels in this application, are particularly useful laboratory tools for the direct visualization of fluid flow revealing pore-scale mechanisms controlling flow and transport phenomena in natural porous media. Current microfluidic devices with uniform etched depths, however, are limited when representing complex geometries such as the multiple-scale pore sizes common in carbonate rocks. In this study, we successfully developed optimized sequential photolithography to etch micropores (1.5 to 21 μm width) less deeply than the depth of wider macropores (＞21 μm width) to improve the structural realism of an existing single-depth micromodel with a carbonate-derived pore structure. Surface profilimetry illustrates the configuration of the dual-depth dual-porosity micromodel and is used to estimate the corresponding pore volume change for the dual-depth micromodel compared to the equivalent uniform- or single-depth model. The flow characteristics of the dual-depth dual-porosity micromodel were characterized using micro-particle image velocimetry (μ-PIV), relative permeability measurements, and pore-scale observations during imbibition and drainage processes. The μ-PIV technique provides insights into the fluid dynamics within microfluidic channels and relevant fluid velocities controlled predominantly by changes in etching depth. In addition, the reduction of end-point relative permeability for both oil and water in the new dual-depth dual-porosity micromodel compared to the equivalent single-depth micromodel implies more realistic capillary forces occurring in the new dual-depth micromodel. Throughout the imbibition and drainage experiments, the flow behaviors of single- and dual-depth micromodels are further differentiated using direct visualization of the trapped non-wetting phase and the preferential mobilization of the wetting phase in the dual-depth micromodel. The visual observations agree with the relative permeability results. These findings indicate that dual-porosity and dual-depth micromodels have enhanced physical realism that is pertinent to oil recovery processes in complex porous media. [ABSTRACT FROM AUTHOR]

Abstract: Oct4–Sox2–Nanog transcriptional networks are critical for the maintenance of embryonic stem (ES) cell self-renewal and induction of pluripotency. However, in transcription factor-induced reprogramming of somatic cells into induced pluripotent stem cells (iPSCs), Nanog is initially dispensable and Oct4 remains the sole factor that could not be substituted/omitted. Here, we show that mouse fibroblasts could be reprogrammed into iPSCs by Nanog and Bmi1, which replaces Sox2, Klf4, and c-Myc, in the absence of Oct4. Furthermore, we show that in the presence of shh agonists (oxysterol and purmophamine), which replaces the function of Bmi1, a single transcription factor, Nanog is sufficient to reprogram mouse fibroblasts into iPSCs. Nanog-induced iPSCs resemble mESCs in terms of morphology, global gene expression profiles, epigenetic status and pluripotency both in vitro and in vivo. These findings support that Nanog can replace the Oct4 for the somatic cell reprogramming and underlie the mechanisms of Nanog in reprogramming process. [Copyright &y& Elsevier]

A procedure combining experiments and deep learning is demonstrated to acquire pore-scale images of oil- and water-wet surfaces over a large field of view in microfluidic devices and to classify wettability based upon these pore scale images. Deep learning supplants the manual, time-consuming, error-prone investigation and categorization of such images. Image datasets were obtained by visualizing the distribution of immiscible phases (n-decane and water) within in-house fabricated micromodels containing sandstone-type and carbonate-type pore structures. The reference dataset consists of 6400 color images binned into four classes for sandstone (water- or oil-wet surfaces) and carbonate (water- or oil-wet surfaces) pore-network patterns. There are 1600 images per class. During 10 sequential training and testing runs of the deep-learning algorithm, 3000, 100, and 100 images were randomly assigned per each rock pattern as the training, validation, and test sets, respectively. We trained and optimized both a Fully Connected Neural Network (FCN) and Convolutional Neural Network (ConvNet) using the image data. The ConvNet performs better as 5 and 8 layers are implemented, as expected. The FCN shows an average test set accuracy for binary surface wettability classification of 87.4% for sandstone rock type and 98.7% for carbonate rock type pore networks. Distinctive heterogeneity in the carbonate rock type and its relevant phase saturation profile resulted in a better prediction accuracy. The best ConvNet models shows an average test set accuracy of binary surface wettability classification of 99.4 ± 0.1% for both sandstone-type and carbonate pore networks. Heterogenous pore sizes and an abundance of small pores amplify the effects of wetting and aid identification. Overall, the test set accuracy for the simultaneous classification of four classes including both sandstone (water- or oil-wet) and carbonate rock pattern (water- or oil-wet) is 98.5% with an 8-layer ConvNet. Performance of the deep-learning model is further interpreted using saliency maps that indicate the degree to which each pixel in the image affects the classification score. Pixels at and adjacent to interfaces are most important to classification. [ABSTRACT FROM AUTHOR]

Abstract Control of adipogenesis in mesenchymal stem cells (MSCs) offers enormous potential for management of obesity- and aging-related diseases. Celastrol, the traditional Chinese medicine extracted from Tripterygium wilfordi , exhibits anti-obesity effects in in vitro and in vivo murine models. This study describes how celastrol affects multilineage differentiation potential of human adipose-derived stem cells (hADSCs). We performed in vitro adipogenic differentiation of hADSCs and investigated how celastrol-induced lipid accumulation and expression of adipocyte differentiation markers varied with dose, duration, and donor age. In addition, we assessed the effect of celastrol on osteogenic and chondrogenic differentiation of hADSCs. During adipogenic induction of hADSCs, the inhibitory effect of celastrol on lipid accumulation and adipogenesis depended on dose, duration, time of administration, and individual donor. Inhibition was mediated by proliferator-activated receptor-γ (PPARG) and CCAAT/enhancer-binding protein alpha (CEBPA). Celastrol also suppressed differentiation of hADSCs into the osteogenic and chondrogenic lineages. Celastrol plays a regulatory role in multilineage differentiation of human MSCs. Our findings provide important insights regarding management of obesity and stem cell therapy. Highlights • Celastrol has therapeutic potential against obesity- and aging-related diseases. • Anti-adipogenic effect of celatrol in human ADSCs depended on dose and duration. • Celatrol inhibited adipogenesis in human ADSCs by regulating PPARG and CEBPA. • Celastrol inhibited adipogenesis in hADSCs derived from 12 donors with various ages. • Celastrol also inhibited osteogenic and chondrogenic differentiation in hADSCs. [ABSTRACT FROM AUTHOR]

Background: Regenerative medicine strategies employing nephron progenitor cells (NPCs) are a viable approach that is worthy of substantial consideration as a promising cell source for kidney diseases. However, the generation of induced nephron progenitor-like cells (iNPCs) from human somatic cells remains a major challenge. Here, we describe a novel method for generating NPCs from human urine-derived cells (UCs) that can undergo long-term expansion in a serum-free condition. Results: Here, we generated iNPCs from human urine-derived cells by forced expression of the transcription factors OCT4, SOX2, KLF4, c-MYC, and SLUG, followed by exposure to a cocktail of defined small molecules. These iNPCs resembled human embryonic stem cell-derived NPCs in terms of their morphology, biological characteristics, differentiation potential, and global gene expression and underwent a long-term expansion in serum-free conditions. Conclusion: This study demonstrates that human iNPCs can be readily generated and expanded, which will facilitate their broad applicability in a rapid, efficient, and patient-specific manner, particularly holding the potential as a transplantable cell source for patients with kidney disease. [ABSTRACT FROM AUTHOR]

The possibility of controlling cell fates by overexpressing specific transcription factors has led to numerous studies in stem cell research. Small molecules can be used, instead of transcription factors, to induce the de-differentiation of somatic cells or to induce pluripotent cells (iPSCs). Here we reported that combinations of small molecules could convert mouse fibroblasts into cardiomyocyte-like cell without requiring transcription factor expression. Treatment with specific combinations of small molecules that are enhancer for iPSC induction converted mouse fibroblasts into spontaneously contracting, cardiac troponin T-positive, cardiomyocyte-like cells. We specifically identified five small molecules that can induce mouse fibroblasts to form these cardiomyocyte-like cells. These cells are similar to primary cardiomyocytes in terms of marker gene expression, epigenetic status of cardiac-specific genes, and subcellular structure. Our findings indicate that lineage conversion can be induced not only by transcription factors, but also by small molecules. [ABSTRACT FROM AUTHOR]

Abstract: Somatic cells can be reprogrammed to generate induced pluripotent stem cells (iPSCs) by overexpression of four transcription factors, Oct4, Klf4, Sox2, and c-Myc. However, exogenous expression of pluripotency factors raised concerns for clinical applications. Here, we show that iPS-like cells (iPSLCs) were generated from mouse somatic cells in two steps with small molecule compounds. In the first step, stable intermediate cells were generated from mouse astrocytes by Bmi1. These cells called induced epiblast stem cell (EpiSC)-like cells (iEpiSCLCs) are similar to EpiSCs in terms of expression of specific markers, epigenetic state, and ability to differentiate into three germ layers. In the second step, treatment with MEK/ERK and GSK3 pathway inhibitors in the presence of leukemia inhibitory factor resulted in conversion of iEpiSCLCs into iPSLCs that were similar to mESCs, suggesting that Bmi1 is sufficient to reprogram astrocytes to partially reprogrammed pluripotency. Next, Bmi1 function was replaced with Shh activators (oxysterol and purmorphamine), which demonstrating that combinations of small molecules can compensate for reprogramming factors and are sufficient to directly reprogram mouse somatic cells into iPSLCs. The chemically induced pluripotent stem cell-like cells (ciPSLCs) showed similar gene expression profiles, epigenetic status, and differentiation potentials to mESCs. [Copyright &y& Elsevier]

Human neural stem cells (NSCs) hold enormous promise for neurological disorders, typically requiring their expandable and differentiable properties for regeneration of damaged neural tissues. Despite the therapeutic potential of induced NSCs (iNSCs), a major challenge for clinical feasibility is the presence of integrated transgenes in the host genome, contributing to the risk for undesired genotoxicity and tumorigenesis. Here, we describe the advanced transgene-free generation of iNSCs from human urine-derived cells (HUCs) by combining a cocktail of defined small molecules with self-replicable mRNA delivery. The established iNSCs were completely transgene-free in their cytosol and genome and further resembled human embryonic stem cell-derived NSCs in the morphology, biological characteristics, global gene expression, and potential to differentiate into functional neurons, astrocytes, and oligodendrocytes. Moreover, iNSC colonies were observed within eight days under optimized conditions, and no teratomas formed in vivo, implying the absence of pluripotent cells. This study proposes an approach to generate transplantable iNSCs that can be broadly applied for neurological disorders in a safe, efficient, and patient-specific manner. [ABSTRACT FROM AUTHOR]