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Collagen high throughput

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https://www.readbyqxmd.com/read/28337850/high-throughput-screening-of-vascular-endothelium-destructive-or-protective-microenvironments-cooperative-actions-of-extracellular-matrix-composition-stiffness-and-structure
#1
Yonghui Ding, Michael Floren, Wei Tan
Pathological modification of the subendothelial extracellular matrix (ECM) has closely been associated with endothelial activation and subsequent cardiovascular disease progression. To understand regulatory mechanisms of these matrix modifications, the majority of previous efforts have focused on the modulation of either chemical composition or matrix stiffness on 2D smooth surfaces without simultaneously probing their cooperative effects on endothelium function on in vivo like 3D fibrous matrices. To this end, a high-throughput, combinatorial microarray platform on 2D and 3D hydrogel settings to resemble the compositions, stiffness, and structure of healthy and diseased subendothelial ECM has been established, and further their respective and combined effects on endothelial attachment, proliferation, inflammation, and junctional integrity have been investigated...
March 24, 2017: Advanced Healthcare Materials
https://www.readbyqxmd.com/read/28322869/scaffold-free-high-throughput-generation-of-quiescent-valvular-microtissues
#2
Annelies Roosens, Inès Puype, Ria Cornelissen
AIMS: The major challenge of working with valvular interstitial cells in vitro is the preservation or recovery of their native quiescent state. In this study, a biomimetic approach is used which aims to engineer small volume, high quality valve microtissues, having a potential in regenerative medicine and as a relevant 3D in vitro model to provide insights into valve (patho)biology. METHODS AND RESULTS: To form micro-aggregates, porcine valvular interstitial cells were seeded in agarose micro-wells and cultured in medium supplemented with 250μM Ascorbic Acid 2-phosphate for 22days...
March 17, 2017: Journal of Molecular and Cellular Cardiology
https://www.readbyqxmd.com/read/28298644/the-combination-of-three-dimensional-and-rotary-cell-culture-system-promotes-the-proliferation-and-maintains-the-differentiation-potential-of-rat-bmscs
#3
Yilong Tang, Yan Xu, Zhifeng Xiao, Yannan Zhao, Jing Li, Sufang Han, Lei Chen, Bin Dai, Ling Wang, Bing Chen, Hong Wang
Bone marrow mesenchymal stem cells (BMSCs) are a good candidate for tissue engineering and clinical application. One of the challenges in its cell therapy is how to quickly obtain an adequate number of seed cells and meanwhile maintain suitable differentiation potential. In this study we combined three-dimensional (3D) collagen porous scaffolds with rotary cell culture system (RCCS) (RCCS-3D) to create a stereoscopic dynamic environment for the amplification of rat BMSCs in vitro. The results revealed that this RCCS-3D system could enhance BMSCs' proliferation and colony formation, as well as maintain the differentiation potential compared with conventional static two-dimensional (2D) and 3D cell culture conditions...
March 15, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28255014/macrothrombocytopenia-and-dense-granule-deficiency-associated-with-fli1-variants-ultrastructural-and-pathogenic-features
#4
Paul Saultier, Léa Vidal, Matthias Canault, Denis Bernot, Céline Falaise, Catherine Pouymayou, Jean-Claude Bordet, Noémie Saut, Agathe Rostan, Véronique Baccini, Franck Peiretti, Marie Favier, Pauline Lucca, Jean-François Deleuze, Robert Olaso, Anne Boland, Pierre Emmanuel Morange, Christian Gachet, Fabrice Malergue, Sixtine Fauré, Anita Eckly, David-Alexandre Trégouët, Marjorie Poggi, Marie-Christine Alessi
Congenital macrothrombocytopenia is a family of rare diseases, of which a significant fraction remains to be genetically characterized. To analyze cases of unexplained thrombocytopenia, 27 individuals from a patient cohort of the Bleeding and Thrombosis Exploration Center of the University Hospital of Marseille were recruited for a high-throughput gene sequencing study. This strategy led to the identification of two novel FLI1 variants (c.1010G>A and c.1033A>G) responsible for macrothrombocytopenia. The FLI1 variant carriers platelets exhibited a defect in aggregation induced by low dose ADP, collagen and TRAP, a defect in ATP secretion, a reduced mepacrine uptake and release and a reduced CD63 expression upon TRAP stimulation...
March 2, 2017: Haematologica
https://www.readbyqxmd.com/read/28240272/a-microfabricated-96-well-3d-assay-enabling-high-throughput-quantification-of-cellular-invasion-capabilities
#5
Rui Hao, Yuanchen Wei, Chaobo Li, Feng Chen, Deyong Chen, Xiaoting Zhao, Shaoliang Luan, Beiyuan Fan, Wei Guo, Junbo Wang, Jian Chen
This paper presents a 96-well microfabricated assay to study three-dimensional (3D) invasion of tumor cells. A 3D cluster of tumor cells was first generated within each well by seeding cells onto a micro-patterned surface consisting of a central fibronectin-coated area that promotes cellular attachment, surrounded by a poly ethylene glycol (PEG) coated area that is resistant to cellular attachment. Following the formation of the 3D cell clusters, a 3D collagen extracellular matrix was formed in each well by thermal-triggered gelation...
February 27, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28216326/a-scalable-lysyl-hydroxylase-2-expression-system-and-luciferase-based-enzymatic-activity-assay
#6
Hou-Fu Guo, Eun Jeong Cho, Ashwini K Devkota, Yulong Chen, William Russell, George N Phillips, Mitsuo Yamauchi, Kevin N Dalby, Jonathan M Kurie
Hydroxylysine aldehyde-derived collagen cross-links (HLCCs) accumulate in fibrotic tissues and certain types of cancer and are thought to drive the progression of these diseases. HLCC formation is initiated by lysyl hydroxylase 2 (LH2), an Fe(II) and α-ketoglutarate (αKG)-dependent oxygenase that hydroxylates telopeptidyl lysine residues on collagen. Development of LH2 antagonists for the treatment of these diseases will require a reliable source of recombinant LH2 protein and a non-radioactive LH2 enzymatic activity assay that is amenable to high throughput screens of small molecule libraries...
March 15, 2017: Archives of Biochemistry and Biophysics
https://www.readbyqxmd.com/read/28207883/application-of-an-optimized-flow-cytometry-based-quantification-of-platelet-activation-pact-monitoring-platelet-activation-in-platelet-concentrates
#7
Cécile H Kicken, Mark Roest, Yvonne M C Henskens, Bas de Laat, Dana Huskens
BACKGROUND: Previous studies have shown that flow cytometry is a reliable test to quantify platelet function in stored platelet concentrates (PC). It is thought that flow cytometry is laborious and hence expensive. We have optimized the flow cytometry-based quantification of agonist induced platelet activation (PACT) to a labor, time and more cost-efficient test. Currently the quality of PCs is only monitored by visual inspection, because available assays are unreliable or too laborious for use in a clinical transfusion laboratory...
2017: PloS One
https://www.readbyqxmd.com/read/28198402/an-integrated-microfluidic-device-for-rapid-and-high-sensitivity-analysis-of-circulating-tumor-cells
#8
Jianing Jiang, Hui Zhao, Weiliang Shu, Jing Tian, Yuqing Huang, Yongxin Song, Ruoyu Wang, Encheng Li, Dennis Slamon, Dongmei Hou, Xiaohui Du, Lichuan Zhang, Yan Chen, Qi Wang
Recently there has been a more focus on the development of an efficient technique for detection of circulating tumor cells (CTCs), due to their significance in prognosis and therapy of metastatic cancer. However, it remains a challenge because of the low count of CTCs in the blood. Herein, a rapid and high-sensitivity approach for CTCs detection using an integrated microfluidic system, consisting of a deterministic lateral displacement (DLD) isolating structure, an automatic purifying device with CD45-labeled immunomagnetic beads and a capturing platform coated with rat-tail collagen was reported...
February 15, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28181140/real-architecture-for-3d-tissue-raft%C3%A2-culture-system-improves-viability-and-maintains-insulin-and-glucagon-production-of-mouse-pancreatic-islet-cells
#9
Gabor J Szebeni, Zsuzsanna Tancos, Liliana Z Feher, Robert Alfoldi, Julianna Kobolak, Andras Dinnyes, Laszlo G Puskas
There is an unmet medical need for the improvement of pancreatic islet maintenance in culture. Due to restricted donor availability it is essential to ameliorate islet viability and graft engraftment. The aim of this study was to compare the standard tissue culture techniques with the advanced Real Architecture For 3D Tissue (RAFT™) culture system in terms of viability and hormone production. Here, we first report that islets embedded in RAFT™ collagen type I advanced tissue culture system maintain their tissue integrity better than in monolayer and suspension cultures...
February 8, 2017: Cytotechnology
https://www.readbyqxmd.com/read/28166248/characterization-of-three-dimensional-cancer-cell-migration-in-mixed-collagen-matrigel-scaffolds-using-microfluidics-and-image-analysis
#10
María Anguiano, Carlos Castilla, Martin Maška, Cristina Ederra, Rafael Peláez, Xabier Morales, Gorka Muñoz-Arrieta, Maite Mujika, Michal Kozubek, Arrate Muñoz-Barrutia, Ana Rouzaut, Sergio Arana, José Manuel Garcia-Aznar, Carlos Ortiz-de-Solorzano
Microfluidic devices are becoming mainstream tools to recapitulate in vitro the behavior of cells and tissues. In this study, we use microfluidic devices filled with hydrogels of mixed collagen-Matrigel composition to study the migration of lung cancer cells under different cancer invasion microenvironments. We present the design of the microfluidic device, characterize the hydrogels morphologically and mechanically and use quantitative image analysis to measure the migration of H1299 lung adenocarcinoma cancer cells in different experimental conditions...
2017: PloS One
https://www.readbyqxmd.com/read/28143748/a-three-dimensional-neural-spheroid-model-for-capillary-like-network-formation
#11
Molly E Boutin, Liana L Kramer, Liane L Livi, Tyler Brown, Christopher Moore, Diane Hoffman-Kim
BACKGROUND: In vitro three-dimensional neural spheroid models have an in vivo-like cell density, and have the potential to reduce animal usage and increase experimental throughput. The aim of this study was to establish a spheroid model to study the formation of capillary-like networks in a three-dimensional environment that incorporates both neuronal and glial cell types, and does not require exogenous vasculogenic growth factors. NEW METHOD: We created self-assembled, scaffold-free cellular spheroids using primary-derived postnatal rodent cortex as a cell source...
January 29, 2017: Journal of Neuroscience Methods
https://www.readbyqxmd.com/read/28129584/bone-natural-autofluorescence-and-confocal-laser-scanning-microscopy-preliminary-results-of-a-novel-useful-tool-to-distinguish-between-forensic-and-ancient-human-skeletal-remains
#12
Luigi Capasso, Ruggero D'Anastasio, Simone Guarnieri, Joan Viciano, Maria Mariggiò
The fast, high-throughput distinction between palaeoanthropological/archaeological remains and recent forensic/clinical bone samples is of vital importance in the field of medico-legal science. In this paper, a novel dating method was developed using the autofluorescence of human bones and the confocal laser scanning microscope as the means to distinguish between archaeological and forensic anthropological skeletal findings. Human bones exhibit fluorescence, typically induced by natural antibiotics that are absorbed by collagen, and provide secondary, exogenous fluorophores...
January 22, 2017: Forensic Science International
https://www.readbyqxmd.com/read/27989921/a-model-system-for-developing-a-tissue-engineered-meniscal-enthesis
#13
Mary Clare McCorry, Melissa M Mansfield, Xiaozhou Sha, Daniel J Coppola, Jonathan W Lee, Lawrence J Bonassar
The meniscus acts as a stabilizer, lubricator, and load distributer in the knee joint. The mechanical stability of the meniscus depends on its connection to the underlying bone by a fibrocartilage to bone transition zone called the meniscal enthesis. Tissue engineered menisci hold great promise as a treatment alternative however lack a means of integrated fixation to the underlying bone needed in order for a tissue engineered meniscal replacement to be successful. Tissue engineering the meniscal enthesis is a difficult task given the complex gradients of cell type, mineral, and extracellular matrix molecules...
October 29, 2016: Acta Biomaterialia
https://www.readbyqxmd.com/read/27870847/creating-high-resolution-multiscale-maps-of-human-tissue-using-multi-beam-sem
#14
André F Pereira, Daniel J Hageman, Tomasz Garbowski, Christof Riedesel, Ulf Knothe, Dirk Zeidler, Melissa L Knothe Tate
Multi-beam scanning electron microscopy (mSEM) enables high-throughput, nano-resolution imaging of macroscopic tissue samples, providing an unprecedented means for structure-function characterization of biological tissues and their cellular inhabitants, seamlessly across multiple length scales. Here we describe computational methods to reconstruct and navigate a multitude of high-resolution mSEM images of the human hip. We calculated cross-correlation shift vectors between overlapping images and used a mass-spring-damper model for optimal global registration...
November 2016: PLoS Computational Biology
https://www.readbyqxmd.com/read/27869911/controlled-production-of-sub-millimeter-liquid-core-hydrogel-capsules-for-parallelized-3d-cell-culture
#15
Hugo Doméjean, Mathieu de la Motte Saint Pierre, Anette Funfak, Nicolas Atrux-Tallau, Kevin Alessandri, Pierre Nassoy, Jérôme Bibette, Nicolas Bremond
Liquid core capsules having a hydrogel membrane are becoming a versatile tool for three-dimensional culture of micro-organisms and mammalian cells. Making sub-millimeter capsules at a high rate, via the breakup of a compound jet in air, opens the way to high-throughput screening applications. However, control of the capsule size monodispersity, especially required for quantitative bioassays, was still lacking. Here, we report how the understanding of the underlying hydrodynamic instabilities that occur during the process can lead to calibrated core-shell bioreactors...
December 20, 2016: Lab on a Chip
https://www.readbyqxmd.com/read/27627810/generation-of-an-in%C3%A2-vitro-3d-pdac-stroma-rich-spheroid-model
#16
Matthew J Ware, Vazrik Keshishian, Justin J Law, Jason C Ho, Carlos A Favela, Paul Rees, Billie Smith, Sayeeduddin Mohammad, Rosa F Hwang, Kimal Rajapakshe, Cristian Coarfa, Shixia Huang, Dean P Edwards, Stuart J Corr, Biana Godin, Steven A Curley
Pancreatic ductal adenocarcinoma (PDAC) is characterized by a prominent desmoplastic/stromal reaction, which contributes to the poor clinical outcome of this disease. Therefore, greater understanding of the stroma development and tumor-stroma interactions is highly required. Pancreatic stellate cells (PSC) are myofibroblast-like cells located in exocrine areas of the pancreas, which as a result of inflammation produced by PDAC migrate and accumulate in the tumor mass, secreting extracellular matrix components and producing the dense PDAC stroma...
November 2016: Biomaterials
https://www.readbyqxmd.com/read/27597161/discovery-of-a-new-structural-class-of-competitive-hdhodh-inhibitors-with-in-vitro-and-in-vivo-anti-inflammatory-immunosuppressive-effects
#17
Wenbin Shen, Xiao Ren, Jingtong Zhu, Yan Xu, Jie Lin, Yeying Li, Feng Zhao, Haizhou Zheng, Ruolan Li, Xiaolan Cui, Xuexia Zhang, Xinhua Lu, Zhihui Zheng
Human dihydroorotate dehydrogenase (hDHODH) is an inner mitochondrial membrane enzyme that involves in the fourth step of the biosynthesis of pyrimidine base. Inhibitors of hDHODH have been proven efficacy for the treatments of inflammation, rheumatoid arthritis, multiple sclerosis and cancer. In the present study, ascochlorin (ASC) and its derivatives, natural compounds from fungal metabolites, were discovered as hDHODH inhibitors by high-throughput screening. Enzyme kinetics studies showed that ASC competitively binds to hDHODH at the site of coenzyme Q substrate...
November 15, 2016: European Journal of Pharmacology
https://www.readbyqxmd.com/read/27564551/full-length-fibronectin-drives-fibroblast-accumulation-at-the-surface-of-collagen-microtissues-during-cell-induced-tissue-morphogenesis
#18
Jasper Foolen, Jau-Ye Shiu, Maria Mitsi, Yang Zhang, Christopher S Chen, Viola Vogel
Generating and maintaining gradients of cell density and extracellular matrix (ECM) components is a prerequisite for the development of functionality of healthy tissue. Therefore, gaining insights into the drivers of spatial organization of cells and the role of ECM during tissue morphogenesis is vital. In a 3D model system of tissue morphogenesis, a fibronectin-FRET sensor recently revealed the existence of two separate fibronectin populations with different conformations in microtissues, i.e. 'compact and adsorbed to collagen' versus 'extended and fibrillar' fibronectin that does not colocalize with the collagen scaffold...
2016: PloS One
https://www.readbyqxmd.com/read/27536780/redifferentiation-of-high-throughput-generated-fibrochondrocyte-micro-aggregates-impact-of-low-oxygen-tension
#19
Elke Berneel, Charlot Philips, Heidi Declercq, Ria Cornelissen
In meniscus tissue engineering strategies, enhancing the matrix quality of the neomeniscal tissue is important. When the differentiated phenotype of fibrochondrocytes is lost, the quality of the matrix becomes compromised. The objective of this study was to produce uniform fibrochondrocyte micro-aggregates with desirable phenotype and tissue homogeneity in large quantities using a simple and reproducible method. Furthermore, we investigated if hypoxia could enhance the matrix quality. Porcine fibrochondrocytes were expanded at 21% oxygen until passage 3 (P3) and a gene expression profile was determined...
2016: Cells, Tissues, Organs
https://www.readbyqxmd.com/read/27498178/combinatorial-extracellular-matrix-microenvironments-promote-survival-and-phenotype-of-human-induced-pluripotent-stem-cell-derived-endothelial-cells-in-hypoxia
#20
Luqia Hou, John Coller, Vanita Natu, Trevor J Hastie, Ngan F Huang
UNLABELLED: Recent developments in cell therapy using human induced pluripotent stem cell-derived endothelial cells (iPSC-ECs) hold great promise for treating ischemic cardiovascular tissues. However, poor post-transplantation viability largely limits the potential of stem cell therapy. Although the extracellular matrix (ECM) has become increasingly recognized as an important cell survival factor, conventional approaches primarily rely on single ECMs for in vivo co-delivery with cells, even though the endothelial basement membrane is comprised of a milieu of different ECMs...
October 15, 2016: Acta Biomaterialia
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