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https://www.readbyqxmd.com/read/28819837/microfluidic-capture-and-multiplex-immunofluorescence-of-circulating-tumor-cells-to-identify-cancer-of-origin
#1
Chian-Hui Lai, Ying-Chih Chang
Circulating tumor cells (CTCs) are an important biomarker and their analysis can be considered a form of "liquid biopsy." The purpose of this book chapter is to describe the use of the 4-channel CMx (cells captured in maximum) microfluidic chip, containing special micropatterns coated with an antibody-conjugated supported lipid bilayer (SLB) on its surface, to capture and isolate CTCs from the blood of cancer patients. Captured CTCs are subsequently released by an air foam to an immunofluorescence (IF) staining panel that enables further analysis, including the identification of the primary cancer source of the CTCs...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28819231/in-vitro-micropatterned-human-pluripotent-stem-cell-test-%C3%A2%C2%B5p-hpst-for-morphometric-based-teratogen-screening
#2
Jiangwa Xing, Yue Cao, Yang Yu, Huan Li, Ziwei Song, Hanry Yu
Exposure to teratogenic chemicals during pregnancy may cause severe birth defects. Due to high inter-species variation of drug responses as well as financial and ethical burdens, despite the widely use of in vivo animal tests, it's crucial to develop highly predictive human pluripotent stem cell (hPSC)-based in vitro assays to identify potential teratogens. Previously we have shown that the morphological disruption of mesoendoderm patterns formed by geometrically-confined cell differentiation and migration using hPSCs could potentially serve as a sensitive morphological marker in teratogen detection...
August 17, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28809821/gradient-strain-chip-for-stimulating-cellular-behaviors-in-cell-laden-hydrogel
#3
Hsin-Yi Hsieh, Chiao-Wen Chu, Ming-Hsuan Chiu, Shueh-Yao Chu, Tsu-Wei Huang, Fan-Gang Tseng
Artificial guidance for cellular alignment is a hot topic in the field of tissue engineering. Most of the previous research has investigated single strain-induced cellular alignment on a cell-laden hydrogel by using complex experimental processes and mass controlling systems, which are usually associated with contamination issues. Thus, in this article, we propose a simple approach to building a gradient static strain using a fluidic chip with a plastic PDMS cover and a UV transparent glass substrate for the stimulation of cellular behavior in a 3D hydrogel...
August 8, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28809529/multiphoton-fabrication-of-fibronectin-functionalized-protein-micropatterns-stiffness-induced-maturation-of-cell-matrix-adhesions-in-human-mesenchymal-stem-cells
#4
Jiaoni Ma, Chuen Wai Li, Nan Huang, Xinna Wang, Minghui Tong, Alfonso H W Ngan, Barbara P Chan
Cell-matrix adhesions are important structures governing the interactions between cells and their microenvironment at the cell-matrix interface. Focal complex (FC) and focal adhesion (FA) have been substantially investigated in conventional 2D culture system using fibroblast as in vitro model. However, the formation of more mature types of cell-matrix adhesion in human mesenchymal stem cells (hMSCs), including fabrillar adhesion (FBA) and 3D matrix adhesion (3DMA), have not been fully elucidated. Here, we try to provide some hints on which niche factor(s) dominate(s) the maturation of FBA and 3DMA by using multiphoton fabrication-based micropatterning technique...
August 15, 2017: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/28803871/an-rnai-screen-in-a-novel-model-of-oriented-divisions-identifies-the-actin-capping-protein-z-%C3%AE-as-an-essential-regulator-of-spindle-orientation
#5
Florencia di Pietro, Léo Valon, Yingbo Li, Rosette Goïame, Auguste Genovesio, Xavier Morin
Oriented cell divisions are controlled by a conserved molecular cascade involving Gαi, LGN, and NuMA. We developed a new cellular model of oriented cell divisions combining micropatterning and localized recruitment of Gαi and performed an RNAi screen for regulators acting downstream of Gαi. Remarkably, this screen revealed a unique subset of dynein regulators as being essential for spindle orientation, shedding light on a core regulatory aspect of oriented divisions. We further analyze the involvement of one novel regulator, the actin-capping protein CAPZB...
August 4, 2017: Current Biology: CB
https://www.readbyqxmd.com/read/28768826/mechanical-signals-activate-p38-mapk-pathway-dependent-reinforcement-of-actin-via-mechanosensitive-hspb1
#6
Laura Hoffman, Christopher C Jensen, Masaaki Yoshigi, Mary Beckerle
Despite the importance of a cell's ability to sense and respond to mechanical force, the molecular mechanisms by which physical cues are converted to cell-instructive chemical information to influence cell behaviors remain to be elucidated. Exposure of cultured fibroblasts to uniaxial cyclic stretch results in an actin stress fiber reinforcement response that stabilizes the actin cytoskeleton. p38 MAPK signaling is activated in response to stretch, and inhibition of p38 MAPK abrogates stretch-induced cytoskeletal reorganization...
August 2, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28766344/cell-shaping-micropatterns-for-quantitative-super-resolution-microscopy-imaging-of-membrane-mechanosensing-proteins
#7
Anthony Fernandez, Markville Bautista, Ramunas Stanciauskas, Taerin Chung, Fabien Pinaud
Patterning cells on microcontact-printed substrates is a powerful approach to control cell morphology and introduce specific mechanical cues on a cell's molecular organization. Although global changes in cellular architectures caused by micropatterns can easily be probed with diffraction-limited optical microscopy, studying molecular reorganizations at the nanoscale demands micropatterned substrates that accommodate the optical requirements of single molecule microscopy techniques. Here, we developed a simple micropatterning strategy that provides control of cellular architectures and is optimized for nanometer accuracy single molecule tracking and three-dimensional super-resolution imaging of plasma and nuclear membrane proteins in cells...
August 9, 2017: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/28761317/physical-disruption-of-cell-cell-contact-induces-vegf-expression-in-rpe-cells
#8
Farhad Farjood, Elizabeth Vargis
PURPOSE: To investigate the role of RPE cell-cell contact in vascular endothelial growth factor (VEGF) protein expression in cultures of primary human RPE (hRPE) cells and a human RPE cell line (ARPE-19). METHODS: Two in vitro methods, scratching and micropatterning, were used to control the physical dissociation of RPE cell-cell junctions. Scratching was performed by scoring monolayers of RPE cells with a cell scraper. Micropatterning was achieved by using a stencil patterning method...
2017: Molecular Vision
https://www.readbyqxmd.com/read/28760810/morphogen-and-community-effects-determine-cell-fates-in-response-to-bmp4-signaling-in-human-embryonic-stem-cells
#9
Anastasiia Nemashkalo, Albert Ruzo, Idse Heemskerk, Aryeh Warmflash
Paracrine signals maintain developmental states and create cell-fate patterns in vivo, and influence differentiation outcomes in human embryonic stem cells (hESCs) in vitro. Systematic investigation of morphogen signaling is hampered by the difficulty of disentangling endogenous signaling from experimentally applied ligands. Here, we grow hESCs in micropatterned colonies of 1-8 cells ("μColonies") to quantitatively investigate paracrine signaling and the response to external stimuli. We examine BMP4-mediated differentiation in μColonies and standard culture conditions and find that in μColonies, above a threshold concentration, BMP4 gives rise to only a single cell fate, contrary to its role as a morphogen in other developmental systems...
July 31, 2017: Development
https://www.readbyqxmd.com/read/28754648/microtopographical-cues-promote-peripheral-nerve-regeneration-via-transient-mtorc2-activation
#10
Suzanne E Thomson, Chloe Charalambous, Carol-Anne Smith, Penelope M Tsimbouri, Theophile Déjardin, Paul J Kingham, Andrew M Hart, Mathis O Riehle
Despite microsurgical repair, recovery of function following peripheral nerve injury is slow and often incomplete. Outcomes could be improved by an increased understanding of the molecular biology of regeneration and by translation of experimental bioengineering strategies. Topographical cues have been shown to be powerful regulators of the rate and directionality of neurite regeneration, and in this study we investigated the downstream molecular effects of linear micropatterned structures in an organotypic explant model...
July 25, 2017: Acta Biomaterialia
https://www.readbyqxmd.com/read/28753740/photopolymerized-micropatterns-with-high-feature-frequencies-overcome-chemorepulsive-borders-to-direct-neurite-growth
#11
Bradley W Tuft, Linjing Xu, Braden Leigh, Daniel Lee, C Allan Guymon, Marlan R Hansen
Developing and regenerating neurites respond to a variety of biophysical and biochemical cues in their microenvironment to reach target cells and establish appropriate synapses. Defining the hierarchal relationship of both types of cues to direct neurite growth carries broad significance for neural development, regeneration, and, in particular, engineering of neural prostheses that improve tissue integration with native neural networks. In this work, chemorepulsive biochemical borders are established on substrates with a range of surface microfeatures to determine the potential of physical cues to overcome conflicting biochemical cues...
July 28, 2017: Journal of Tissue Engineering and Regenerative Medicine
https://www.readbyqxmd.com/read/28750339/differential-neuronal-and-glial-behavior-on-flat-and-micro-patterned-chitosan-films
#12
Marta Mattotti, Zaida Alvarez, Luis Delgado, Miguel A Mateos-Timoneda, Conrado Aparicio, Josep A Planell, Soledad Alcántara, Elisabeth Engel
Chitosan is a biodegradable natural polysaccharide that has been widely studied for regenerative purposes in the central nervous system. In this study we assessed the in vitro glial and neuronal cells response to chitosan either flat or patterned with grooves in the micrometric range. Chitosan demonstrated to be a good substrate for the attachment and growth of both neurons and glial cells. Chitosan micropatterns promoted glial cell maturation, suggesting astroglial activation. Nevertheless, those mature/reactive glial cells were permissive for axonal growth...
July 19, 2017: Colloids and Surfaces. B, Biointerfaces
https://www.readbyqxmd.com/read/28746386/tracmit-an-effective-pipeline-for-tracking-and-analyzing-cells-on-micropatterns-through-mitosis
#13
Olivier Burri, Benita Wolf, Arne Seitz, Pierre Gönczy
The use of micropatterns has transformed investigations of dynamic biological processes by enabling the reproducible analysis of live cells using time-lapse fluorescence microscopy. With micropatterns, thousands of individual cells can be efficiently imaged in parallel, rendering the approach well suited for screening projects. Despite being powerful, such screens remain challenging in terms of data handling and analysis. Typically, only a fraction of micropatterns is occupied in a manner suitable to monitor a given phenotypic output...
2017: PloS One
https://www.readbyqxmd.com/read/28732996/nanostructured-%C3%AE-type-titanium-alloy-fabricated-by-ultrasonic-nanocrystal-surface-modification
#14
Mehdi Kheradmandfard, Seyed Farshid Kashani-Bozorg, Chang-Lae Kim, Abbas Zarei Hanzaki, Young-Shik Pyoun, Jung-Hyong Kim, Auezhan Amanov, Dae-Eun Kim
The surface of β-type Ti-Nb-Ta-Zr (TNTZ) alloy, which is a promising material for biomedical applications, was treated with the ultrasonic nanocrystal surface modification (UNSM) technique to enhance its hardness. As a result, a gradient nanostructured (GNS) layer was generated in the surface; the microstructure of the top surface layer consisted of nanoscale lamellae with a width of about 60-200nm. In addition, there were lamellar grains consisting of nanostructured subgrains having unclear and wavy boundaries...
November 2017: Ultrasonics Sonochemistry
https://www.readbyqxmd.com/read/28726917/engineering-micromyocardium-to-delineate-cellular-and-extracellular-regulation-of-myocardial-tissue-contractility
#15
Nethika R Ariyasinghe, Caitlin H Reck, Alyssa A Viscio, Andrew P Petersen, Davi M Lyra-Leite, Nathan Cho, Megan L McCain
Cardiovascular diseases are a leading cause of death, in part due to limitations of existing models of the myocardium. Myocardium consists of aligned, contractile cardiac myocytes interspersed with fibroblasts that synthesize extracellular matrix (ECM). The cellular demographics and biochemical and mechanical properties of the ECM remodel in many different cardiac diseases. However, the impact of diverse cellular and extracellular remodeling on the contractile output of the myocardium are poorly understood...
July 20, 2017: Integrative Biology: Quantitative Biosciences From Nano to Macro
https://www.readbyqxmd.com/read/28722147/surface-isoelectric-focusing-sief-with-carrier-ampholyte-ph-gradient
#16
Zhichao Wang, Cornelius Ivory, Adrienne R Minerick
Isoelectric focusing (IEF) is a powerful tool for amphoteric protein separations because of high sensitivity, bio-compatibility, and reduced complexity compared to chromatography or mechanical separation techniques. IEF miniaturization is attractive because it enables rapid analysis, easier adaptation to point of care applications, and smaller sample demands. However, existing small-scale IEF tools have not yet been able to analyze single protein spots from array libraries, which are ubiquitous in many pharmaceutical discovery and screening protocols...
July 19, 2017: Electrophoresis
https://www.readbyqxmd.com/read/28722137/precise-and-arbitrary-deposition-of-biomolecules-onto-biomimetic-fibrous-matrices-for-spatially-controlled-cell-distribution-and-functions
#17
Chao Jia, Bowen Luo, Haoyu Wang, Yongqian Bian, Xueyong Li, Shaohua Li, Hongjun Wang
Advances in nano-/microfabrication allow the fabrication of biomimetic substrates for various biomedical applications. In particular, it would be beneficial to control the distribution of cells and relevant biomolecules on an extracellular matrix (ECM)-like substrate with arbitrary micropatterns. In this regard, the possibilities of patterning biomolecules and cells on nanofibrous matrices are explored here by combining inkjet printing and electrospinning. Upon investigation of key parameters for patterning accuracy and reproducibility, three independent studies are performed to demonstrate the potential of this platform for: i) transforming growth factor (TGF)-β1-induced spatial differentiation of fibroblasts, ii) spatiotemporal interactions between breast cancer cells and stromal cells, and iii) cancer-regulated angiogenesis...
July 19, 2017: Advanced Materials
https://www.readbyqxmd.com/read/28720761/parylene-c-topographic-micropattern-as-a-template-for-patterning-pdms-and-polyacrylamide-hydrogel
#18
Ilaria Sanzari, Mauro Callisti, Antonio De Grazia, Daniel J Evans, Tomas Polcar, Themistoklis Prodromakis
Parylene C is a well-known polymer and it has been mainly employed as a protective layer for implantable electronics. In this paper, we propose a new approach to use Parylene C as a versatile template for patterning soft materials potentially applicable as scaffolds in cardiac tissue engineering (TE). Parylene C substrates were anisotropically patterned through standard lithographic process with hydrophilic channels separating raised hydrophobic strips. Ridges and grooves of the template are 10 µm width and depth ranging from 1 to 17 µm...
July 18, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28707348/regulation-of-cell-arrangement-using-a-novel-composite-micropattern
#19
Xiaoyi Liu, Yaoping Liu, Feng Zhao, Tingting Hun, Shan Li, Yuguang Wang, Weijie Sun, Wei Wang, Yan Sun, Yubo Fan
Micropatterning technique has been used to control single cell geometry in many researches, however, this is no report that it is used to control multicelluar geometry, which not only control single cell geometry but also organize those cells by a certain pattern. In this work, a composite protein micropattern is developed to control both cell shape and cell location simultaneously. The composite micropattern consists of a central circle 15 μm in diameter for single-cell capture, surrounded by small, square arrays (3 μm × 3 μm) for cell spreading...
July 13, 2017: Journal of Biomedical Materials Research. Part A
https://www.readbyqxmd.com/read/28702667/microengineered-cultures-containing-human-hepatic-stellate-cells-and-hepatocytes-for-drug-development
#20
Matthew D Davidson, David A Kukla, Salman R Khetani
In non-alcoholic steatohepatitis (NASH), hepatic stellate cells (HSC) differentiate into myofibroblast-like cells that cause fibrosis, which predisposes patients to cirrhosis and hepatocellular carcinoma. Thus, modeling interactions between activated HSCs and hepatocytes in vitro can aid in the development of anti-NASH/fibrosis therapeutics and lead to a better understanding of disease progression. Species-specific differences in drug metabolism and disease pathways now necessitate the supplementation of animal studies with data acquired using human liver models; however, current models do not adequately model the negative effects of primary human activated HSCs on the phenotype of otherwise well-differentiated primary human hepatocytes (PHHs) as in vivo...
August 14, 2017: Integrative Biology: Quantitative Biosciences From Nano to Macro
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