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https://www.readbyqxmd.com/read/29227553/fluorescence-lifetime-imaging-of-a-caspase-3-apoptosis-reporter
#1
Johanna M Buschhaus, Anne E Gibbons, Kathryn E Luker, Gary D Luker
Caspase-3 is a proteolytic enzyme that functions as a key effector in apoptotic cell death. Determining activity of caspase-3 provides critical information about cancer cell viability and response to treatment. To measure apoptosis in intact cells and living mice, a fluorescence imaging reporter that detects caspase-3 activity by Förster resonance energy transfer (FRET) was used. Changes in FRET by fluorescence lifetime imaging microscopy (FLIM) were measured. Unlike FRET measurements based on fluorescence intensity, lifetime measurements are independent of reporter concentration and scattering of light in tissue, making FLIM a robust method for imaging in 3D environments...
December 11, 2017: Current Protocols in Cell Biology
https://www.readbyqxmd.com/read/29227036/bottom-up-nanoencapsulation-from-single-cells-to-tunable-and-scalable-cellular-spheroids-for-hair-follicle-regeneration
#2
Jin Wang, Yong Miao, Yong Huang, Bojie Lin, Xiaomin Liu, Shune Xiao, Lijuan Du, Zhiqi Hu, Malcolm Xing
Cell surface engineering technology advances cell therapeutics and tissue engineering by accurate micro/nanoscale control in cell-biomaterial ensembles and cell spheroids formation. By tailoring cell surface, microgels can encapsulate cells for versatile uses. However, microgels are coated in a thick layer to house multiple cells together but not a single cell based. Besides, excessive deposition on cell surface is detrimental to cellular functions. Herein, layer-by-layer (LbL) self-assembly to encapsulate single cell using nanogel is reported, owing to its security and tunable thickness at nanoscale, and further forms cell spheroids by physical cross-linking on nanogel-coated cells for delivery...
December 11, 2017: Advanced Healthcare Materials
https://www.readbyqxmd.com/read/29226086/fibroblasts-promote-the-collective-invasion-of-ameloblastoma-tumor-cells-in-a-3d-coculture-model
#3
Takao Fuchigami, Hirofumi Koyama, Michiko Kishida, Yoshiaki Nishizawa, Mikio Iijima, Toshiro Kibe, Masahiro Ueda, Tohru Kiyono, Yoshimasa Maniwa, Norifumi Nakamura, Shosei Kishida
Ameloblastoma is a benign tumor of the odontogenic epithelium with several histological subtypes. All subtypes of ameloblastoma contain abundant stroma; the tumor cells invade collectively into the surrounding tissues without losing intratumor cell attachments. However, the molecular mechanisms mediating ameloblastoma invasion remain unclear. Here, we evaluated the functional significance of the interactions between ameloblastoma tumor cells and stromal fibroblasts on collective cellular invasion using a three-dimensional cultivation method, double-layered collagen gel hemisphere (DL-CGH) culture...
December 2017: FEBS Open Bio
https://www.readbyqxmd.com/read/29223705/functional-hepatocyte-clusters-on-bioactive-blend-silk-matrices-towards-generating-bioartificial-liver-constructs
#4
Guru Janani, Samit K Nandi, Biman B Mandal
The creation of in vitro functional hepatic tissue simulating micro-environmental niche of native liver is a keen area of research due to its demand in bioartificial liver (BAL) and cell-based tissue engineering. Here, we investigated the potential of novel blend (BA) silk scaffold fabricated by blending mulberry (Bombyx mori, BM) silk fibroin with cell adhesion motif (RGD) rich non-mulberry (Antheraea assamensis, AA) silk fibroin in generating a functional liver construct. Three-dimensional (3D) porous silk scaffolds (BM, AA and BA) were physico-chemically characterized and functionally evaluated using human hepatocarcinoma cells (HepG2) and primary neonatal rat hepatocytes...
December 6, 2017: Acta Biomaterialia
https://www.readbyqxmd.com/read/29223312/bioprinting-for-neural-tissue-engineering
#5
REVIEW
Stephanie Knowlton, Shivesh Anand, Twisha Shah, Savas Tasoglu
Bioprinting is a method by which a cell-encapsulating bioink is patterned to create complex tissue architectures. Given the potential impact of this technology on neural research, we review the current state-of-the-art approaches for bioprinting neural tissues. While 2D neural cultures are ubiquitous for studying neural cells, 3D cultures can more accurately replicate the microenvironment of neural tissues. By bioprinting neuronal constructs, one can precisely control the microenvironment by specifically formulating the bioink for neural tissues, and by spatially patterning cell types and scaffold properties in three dimensions...
December 6, 2017: Trends in Neurosciences
https://www.readbyqxmd.com/read/29222846/influence-of-the-three-dimensional-culture-of-human-bone-marrow-mesenchymal-stromal-cells-within-a-macroporous-polysaccharides-scaffold-on-pannexin-1-and-pannexin-3
#6
Julien Guerrero, Hugo Oliveira, Rachida Aid, Reine Bareille, Robin Siadous, Didier Letourneur, Yong Mao, Joachim Kohn, Joëlle Amédée
Because cell interactions play a fundamental role for cell differentiation, we investigated the expression of Pannexin 1 and Pannexin 3 in human bone marrow mesenchymal stromal cells (HBMSCs) in a three-dimensional (3D) microenvironment provided by a polysaccharide-based macroporous scaffold. The pannexin (Panx) family consists of three members, Panx1, Panx2, and Panx3. The roles of pannexin large-pore ion and metabolite channels are recognized in many physiological and pathophysiological scenarios but the role of these proteins in human physiological processes is still under investigation...
December 9, 2017: Journal of Tissue Engineering and Regenerative Medicine
https://www.readbyqxmd.com/read/29219039/morphogenesis-and-mucus-production-of-epithelial-tissues-of-three-major-salivary-glands-of-embryonic-mouse-in-3d-culture
#7
Azusa Nakao, Takumi Inaba, Akiko Murakami-Sekimata, Hiroyuki Nogawa
Embryonic mouse submandibular epithelia initiate branching morphogenesis within two days when embedded in Matrigel and stimulated by members of the epidermal growth factor family. However, it is unknown whether the end buds further branch over longer culture periods, and whether saliva-producing cells differentiate there. In the present study, we cultivated three major (submandibular, sublingual and parotid) salivary epithelia from 13-day embryos for 14 days in mesenchyme-free cultures. All epithelia continued to grow and branch to form numerous acinus-like structures in medium supplemented with neuregulin 1, fibroblast growth factor 1, and insulintransferrin-sodium selenite...
December 2017: Zoological Science
https://www.readbyqxmd.com/read/29216013/bio-inspired-synthesis-of-a-hierarchical-self-assembled-zinc-phosphate-nanostructure-in-the-presence-of-cowpea-mosaic-virus-in-vitro-cell-cycle-proliferation-and-prospects-for-tissue-regeneration
#8
Koushi Kumar, Vasudevarao Penugurti, Gila Levi, Yitzhak Mastai, Bramanandam Manavathi, Pradip Paik
Self-assembly is an important auto-organization process used in designing structural biomaterials which have the potential capability to heal tissues after traumatic injury. Although various materials having the ability to heal after injury are available, there is still a substantial need to develop new improved materials. To address this issue, we have developed hierarchical three-dimensional (3D) self-assembled zinc phosphate (Zn3(PO4)2) in the presence of cowpea mosaic virus (CPMV). Zn3(PO4)2 nanoparticles are self-assembled into nanosheets with a high degree of isotropy and then self-organized into a 3D structure that can enhance surface interactions with biological entities...
December 7, 2017: Biomedical Materials
https://www.readbyqxmd.com/read/29214365/profiling-differential-microrna-expression-between-in-situ-infiltrative-and-lympho-vascular-space-invasive-breast-cancer-a-pilot-study
#9
Patsy S Soon, Pamela J Provan, Edward Kim, Nirmala Pathmanathan, Dinny Graham, Christine L Clarke, Rosemary L Balleine
Ductal carcinoma in situ (DCIS), invasive breast cancer (IBC) and lympho-vascular invasion (LVI) represent distinct stages in breast cancer progression with different clinical implications. Altered microRNA (miRNA) expression may play a role in mediating the progression of DCIS to IBC and LVI. The aim of this pilot study was to investigate whether differential miRNA expression could play a role in breast cancer progression. Cancer cells from DCIS, IBC and LVI were microdissected from formalin fixed paraffin embedded (FFPE) tissue of five breast cancer samples...
December 6, 2017: Clinical & Experimental Metastasis
https://www.readbyqxmd.com/read/29208553/fibril-growth-kinetics-link-buffer-conditions-and-topology-of-3d-collagen-i-networks
#10
Liv Kalbitzer, Tilo Pompe
Three-dimensional fibrillar networks reconstituted from collagen I are widely used as biomimetic scaffolds for in vitro and in vivo cell studies. Various physicochemical parameters of buffer conditions for in vitro fibril formation are well known, including pH-value, ion concentrations and temperature. However, there is a lack of a detailed understanding of reconstituting well-defined 3D network topologies, which is required to mimic specific properties of the native extracellular matrix. We screened a wide range of relevant physicochemical buffer conditions and characterized the topology of the reconstituted 3D networks in terms of mean pore size and fibril diameter...
December 2, 2017: Acta Biomaterialia
https://www.readbyqxmd.com/read/29208552/controlling-human-corneal-stromal-stem-cell-contraction-to-mediate-rapid-cell-and-matrix-organization-of-real-architecture-for-3-dimensional-tissue-equivalents
#11
Dev Mukhey, James Phillips, Julie T Daniels, Alvena K Kureshi
The architecture of the human corneal stroma consists of a highly organized extracellular matrix (ECM) interspersed with keratocytes. Their progenitor cells; corneal stromal stem cells (CSSC) are located at the periphery, in the limbal stroma. A highly organized corneal ECM is critical for effective transmission of light but this structure may be compromised during injury or disease, resulting in loss of vision. Re-creating normal organization in engineered tissue equivalents for transplantation often involves lengthy culture times that are inappropriate for clinical use or utilisation of synthetic substrates that bring complications such as corneal melting...
December 2, 2017: Acta Biomaterialia
https://www.readbyqxmd.com/read/29208458/inhibition-of-tgf-%C3%AE-pathway-reverts-extracellular-matrix-remodeling-in-t-cruzi-infected-cardiac-spheroids
#12
Patrícia M Ferrão, Líndice M Nisimura, Otacílio C Moreira, Marcelo G Land, Mirian C Pereira, Leila de Mendonça-Lima, Tania C Araujo-Jorge, Mariana C Waghabi, Luciana R Garzoni
Chagasic cardiomyopathy (CC) is the main manifestation of Chagas Disease (CD). CC is a progressive dysfunctional illness, in which transforming growth factor beta (TGF-β) plays a central role in fibrogenesis and hypertrophy. In the present study, we tested in a three-dimensional (3D) model of cardiac cells culture (named cardiac spheroids), capable of mimicking the aspects of fibrosis and hypertrophy observed in CC, the role of TGF-β pathway inhibition in restoring extracellular matrix (ECM) balance disrupted by T...
December 2, 2017: Experimental Cell Research
https://www.readbyqxmd.com/read/29205905/a-simple-modification-method-to-obtain-anisotropic-and-porous-3d-microfibrillar-scaffolds-for-surgical-and-biomedical-applications
#13
Vahid Hosseini, Olivera Evrova, Simon P Hoerstrup, Viola Vogel
In native tissues, cellular organization is predominantly anisotropic. Yet, it remains a challenge to engineer anisotropic scaffolds that promote anisotropic cellular organization at macroscopic length scales. To overcome this challenge, an innovative, cheap and easy method to align clinically approved non-woven surgical microfibrillar scaffolds is presented. The method involves a three-step process of coating, unidirectional stretching of scaffolds after heating them above glass transition temperature, and cooling back to room temperature...
December 4, 2017: Small
https://www.readbyqxmd.com/read/29201905/evaluation-of-resins-for-stereolithographic-3d-printed-surgical-guides-the-response-of-l929-cells-and-human-gingival-fibroblasts
#14
Christoph Kurzmann, Klara Janjić, Hassan Shokoohi-Tabrizi, Michael Edelmayer, Manuela Pensch, Andreas Moritz, Hermann Agis
Additive manufacturing is becoming increasingly important in dentistry for the production of surgical guides. The development of cost-effective desktop stereolithography (SLA) printing systems and the corresponding resins makes this novel technique accessible to dental offices and dental laboratories. The aim of the study was to reveal the response of soft tissue cells to Clear and Dental SG resins used in desktop SLA printing systems at different stages of processing. Cell activity of L929 cells and gingival fibroblasts (GF) in response to the materials was examined in indirect and direct monolayer culture models and a direct spheroid culture model based on MTT, resazurin-based toxicity assays, and live-dead staining...
2017: BioMed Research International
https://www.readbyqxmd.com/read/29198564/linking-stem-cell-function-and-growth-pattern-of-intestinal-organoids
#15
Torsten Thalheim, Marianne Quaas, Maria Herberg, Ulf-Dietrich Braumann, Christiane Kerner, Markus Loeffler, Gabriela Aust, Joerg Galle
Intestinal stem cells (ISCs) require well-defined signals from their environment in order to carry out their specific functions. Most of these signals are provided by neighboring cells that form a stem cell niche, whose shape and cellular composition self-organize. Major features of this self-organization can be studied in ISC-derived organoid culture. In this system, manipulation of essential pathways of stem cell maintenance and differentiation results in well-described growth phenotypes. We here provide an individual cell-based model of intestinal organoids that enables a mechanistic explanation of the observed growth phenotypes...
October 31, 2017: Developmental Biology
https://www.readbyqxmd.com/read/29196977/self-organized-cerebellar-tissue-from-human-pluripotent-stem-cells-and-disease-modeling-with-patient-derived-ipscs
#16
REVIEW
Keiko Muguruma
Recent advances in the techniques that differentiate induced pluripotent stem cells (iPSCs) into specific types of cells enabled us to establish in vitro cell-based models as a platform for drug discovery. iPSC-derived disease models are advantageous to generation of a large number of cells required for high-throughput screening. Furthermore, disease-relevant cells differentiated from patient-derived iPSCs are expected to recapitulate the disorder-specific pathogenesis and physiology in vitro. Such disease-relevant cells will be useful for developing effective therapies...
December 2, 2017: Cerebellum
https://www.readbyqxmd.com/read/29192297/rapid-spheroid-clearing-on-a-microfluidic-chip
#17
Tomas Silva Santisteban, Omid Rabajania, Iana Kalinina, Stephen Robinson, Matthias Meier
Spheroids are three-dimensional (3D) cell cultures that aim to bridge the gap between the use of whole animals and cellular monolayers. Microfluidics is regarded as an enabling technology to actively control the chemical environment of 3D cell cultures. Although a wide variety of platforms have been developed to handle spheroid cultures, the development of analytical systems for spheroids remains a major challenge. In this study, we engineered a microfluidic large-scale integration (mLSI) chip platform for tissue-clearing and imaging...
December 1, 2017: Lab on a Chip
https://www.readbyqxmd.com/read/29192263/comprehensive-high-throughput-image-analysis-for-therapeutic-efficacy-of-architecturally-complex-heterotypic-organoids
#18
Anne-Laure Bulin, Mans Broekgaarden, Tayyaba Hasan
Bioengineered three-dimensional (3D) tumor models that incorporate heterotypic cellular communication are gaining interest as they can recapitulate key features regarding the intrinsic heterogeneity of cancer tissues. However, the architectural complexity and heterogeneous contents associated with these models pose a challenge for toxicological assays to accurately report treatment outcomes. To address this issue, we describe a comprehensive image analysis procedure for structurally complex organotypic cultures (CALYPSO) applied to fluorescence-based assays to extract multiparametric readouts of treatment effects for heterotypic tumor cultures that enables advanced analyses...
November 30, 2017: Scientific Reports
https://www.readbyqxmd.com/read/29191717/improved-in-vitro-models-for-preclinical-drug-and-formulation-screening-focusing-on-2d-and-3d-skin-and-cornea-constructs
#19
REVIEW
Nicole Beißner, Antonio Bolea Albero, Jendrik Füller, Thomas Kellner, Lothar Lauterboeck, Jinghu Liang, Markus Böl, Birgit Glasmacher, Christel C Müller-Goymann, Stephan Reichl
The present overview deals with current approaches for the improvement of in vitro models for preclinical drug and formulation screening which were elaborated in a joint project at the Center of Pharmaceutical Engineering of the TU Braunschweig. Within this project a special focus was laid on the enhancement of skin and cornea models. For this reason, first, a computation-based approach for in silico modeling of dermal cell proliferation and differentiation was developed. The simulation should for example enhance the understanding of the performed 2D in vitro tests on the antiproliferative effect of hyperforin...
November 27, 2017: European Journal of Pharmaceutics and Biopharmaceutics
https://www.readbyqxmd.com/read/29191507/viscoelastic-properties-of-human-pancreatic-tumors-and-in-vitro-constructs-to-mimic-mechanical-properties
#20
Andres Rubiano, Daniel Delitto, Song Han, Michael Gerber, Carly Galitz, Jose Trevino, Ryan M Thomas, Steven J Hughes, Chelsey S Simmons
Pancreatic ductal adenocarcinoma (PDAC) is almost universally fatal, in large part due to a protective fibrotic barrier generated by tumor-associated stromal (TAS) cells. This barrier is thought to promote cancer cell survival and confounds attempts to develop effective therapies. We present a 3D in vitro system that replicates the mechanical properties of the PDAC microenvironment, representing an invaluable tool for understanding the biology of the disease. Mesoscale indentation quantified viscoelastic metrics of resected malignant tumors, inflamed chronic pancreatitis regions, and histologically normal tissue...
November 27, 2017: Acta Biomaterialia
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