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organotypic culture

Julien Muffat, Yun Li, Rudolf Jaenisch
In vitro differentiation of human pluripotent stem cells provides a systematic platform to investigate the physiological development and function of the human nervous system, as well as the etiology and consequence when these processes go awry. Recent development in three-dimensional (3D) organotypic culture systems allows modeling of the complex structure formation of the human CNS, and the intricate interactions between various resident neuronal and glial cell types. Combined with an ever-expanding genome editing and regulation toolkit such as CRISPR/Cas9, it is now a possibility to study human neurological disease in the relevant molecular, cellular and anatomical context...
October 18, 2016: Current Opinion in Cell Biology
D K Rieger, E Navarro, I Buendia, E Parada, L González-Lafuente, R Leon, A P Costa, I A Heinrich, K S Nascimento, B S Cavada, M G Lopez, J Egea, R B Leal
Lectins are proteins that bind cellular glycans and can modulate various neuronal functions. We have evaluated the neuroprotective effect of ConBr, a lectin purified from the seeds of Canavalia brasiliensis in a model of rat organotypic hippocampal cultures (OHCs) exposed to oxygen and glucose deprivation (OGD). OGD for 15 min followed by 24 h re-oxygenation significantly increased cell death, caused mitochondrial depolarization and increased reactive oxygen species (ROS) in CA1 region of OHCs. ConBr (0.1 μg/mL) added during the re-oxygenation period counteracted cell death, mitochondrial depolarization and overproduction of ROS induced by OGD...
October 17, 2016: Neurochemical Research
Tuula M Eriksson, Richard M Day, Stefano Fedele, Vehid M Salih
Ameloblastoma is a rare, odontogenic neoplasm with benign histopathology, but extensive, local infiltrative capacity through the bone tissue it originates in. While the mechanisms of ameloblastoma invasion through the bone and bone absorption are largely unknown, recent investigations have indicated a role of the osteoprotegerin/receptor activator of nuclear factor kappa-B ligand regulatory mechanisms. Here, we present results obtained using a novel in vitro organotypic tumour model, which we have developed using tissue engineering techniques...
January 2016: Journal of Tissue Engineering
Ana Gutiérrez-Franco, Herena Eixarch, Carme Costa, Vanessa Gil, Mireia Castillo, Laura Calvo-Barreiro, Xavier Montalban, José A Del Río, Carmen Espejo
Semaphorin 7A (sema7A) is classified as an immune semaphorin with dual functions in the immune system and in the central nervous system (CNS). These molecules are of interest due to their potential role in multiple sclerosis (MS), which is a chronic demyelinating and neurodegenerative disease of autoimmune origin. In this study, we elucidated the role of sema7A in neuroinflammation using both in vitro and in vivo experimental models. In an in vitro model of neuroinflammation, using cerebellar organotypic slice cultures, we observed that challenge with lipopolysaccharide (LPS) endotoxin did not affect demyelination or cell death in sema7A-deficient cultures compared to wild-type cultures...
October 6, 2016: Molecular Neurobiology
Anne Tscherter, Martina Heidemann, Sonja Kleinlogel, Jürg Streit
Presently there exists no cure for spinal cord injury (SCI). However, transplantation of embryonic tissue into spinal cord (SC) lesions resulted in axon outgrowth across the lesion site and some functional recovery, fostering hope for future stem cell therapies. Although in vivo evidence for functional recovery is given, the exact cellular mechanism of the graft support remains elusive: either the grafted cells provide a permissive environment for the host tissue to regenerate itself or the grafts actually integrate functionally into the host neuronal network reconnecting the separated SC circuits...
2016: Frontiers in Cellular Neuroscience
Charlie J Gilbride
Organotypic hippocampal slice cultures (OHSCs) provide the experimental flexibility of cell culture while leaving much of the natural neuronal connectivity intact. Previously, it was shown that the functional and morphological features of CA1 pyramidal neurons in OHSCs resemble, to a surprising extent, those of CA1 neurons in the acute brain slice preparation. However, the extent to which the characteristics of other principle hippocampal neurons change or are preserved in cultured slices remains to be determined...
October 2016: Physiological Reports
Jacob I Ayers, Jeffrey Diamond, Adriana Sari, Susan Fromholt, Ahmad Galaleldeen, Lyle W Ostrow, Jonathan D Glass, P John Hart, David R Borchelt
Evidence of misfolded wild-type superoxide dismutase 1 (SOD1) has been detected in spinal cords of sporadic ALS (sALS) patients, suggesting an etiological relationship to SOD1-associated familial ALS (fALS). Given that there are currently a number of promising therapies under development that target SOD1, it is of critical importance to better understand the role of misfolded SOD1 in sALS. We previously demonstrated the permissiveness of the G85R-SOD1:YFP mouse model for MND induction following injection with tissue homogenates from paralyzed transgenic mice expressing SOD1 mutations...
October 4, 2016: Acta Neuropathologica
Elizabeth A Calle, Ryan C Hill, Katherine L Leiby, Andrew V Le, Ashley L Gard, Joseph A Madri, Kirk C Hansen, Laura E Niklason
: Extracellular matrix is a key component of many products in regenerative medicine. Multiple regenerative medicine products currently in the clinic are comprised of human or xenogeneic extracellular matrix. In addition, whole-organ regeneration exploits decellularized native organs as scaffolds for organotypic cell culture. However, precise understanding of the constituents of such extracellular matrix-based implants and scaffolds has sorely lagged behind their use. We present here an advanced protein extraction method using known quantities of proteotypic (13)C-labeled peptides to quantify matrix proteins in native and decellularized lung tissues...
September 29, 2016: Acta Biomaterialia
Karl Frontzek, Manuela Pfammatter, Silvia Sorce, Assunta Senatore, Petra Schwarz, Rita Moos, Katrin Frauenknecht, Simone Hornemann, Adriano Aguzzi
Prions are the infectious agents causing transmissible spongiform encephalopathies (TSE), progressive, inexorably lethal neurological diseases. Antibodies targeting the globular domain (GD) of the cellular prion protein PrPC trigger a neurotoxic syndrome morphologically and molecularly similar to prion disease. This phenomenon raises the question whether such antibodies induce infectious prions de novo. Here we exposed cerebellar organotypic cultured slices (COCS) to the neurotoxic antibody, POM1. We then inoculated COCS homogenates into tga20 mice, which overexpress PrPC and are commonly utilized as sensitive indicators of prion infectivity...
2016: PloS One
Enzo Wanke, Francesca Gullo, Elena Dossi, Gaetano Valenza, Andrea Becchetti
Astrocytes reuptake synaptically released glutamate with electrogenic transporters (GluT), and buffer the spike-dependent extracellular K(+) ([K(+)]o) excess with background K(+) channels. We studied neuronal spikes and the slower astrocytic signals on reverberating neocortical cultures and organotypic slices from mouse brains. Spike trains and glial responses were simultaneously captured from individual sites of multi-electrode arrays (MEA), by splitting the recorded traces into appropriate filters, and reconstructing the original signal by deconvolution...
September 28, 2016: Journal of Neurophysiology
Ekaterina V Lopatina, Anna V Kipenko, Natalia A Pasatetskaya, Valentina A Penniyaynen, Boris V Krylov
Endogenous digitalis-like factors were found in the mammalian and human blood. It was the starting point for the elucidation of the new non-pumping function of the Na(+),K(+)-ATPase. It was previously well known that Na(+),K(+)-ATPase is a pharmacological target receptor for cardiac glycosides (J.C. Skou. 1957. Biochim. Biophys. Acta, 23: 394-401). We have investigated the trophotropic effects of such agents as ouabain, epinephrine, norepinephrine, atenolol, and comenic acid using the organotypic tissue culture combined with the reconstruction of optical cross sections and confocal microscopy...
October 2016: Canadian Journal of Physiology and Pharmacology
Julien Muffat, Yun Li, Bingbing Yuan, Maisam Mitalipova, Attya Omer, Sean Corcoran, Grisilda Bakiasi, Li-Huei Tsai, Patrick Aubourg, Richard M Ransohoff, Rudolf Jaenisch
Microglia, the only lifelong resident immune cells of the central nervous system (CNS), are highly specialized macrophages that have been recognized to have a crucial role in neurodegenerative diseases such as Alzheimer's, Parkinson's and adrenoleukodystrophy (ALD). However, in contrast to other cell types of the human CNS, bona fide microglia have not yet been derived from cultured human pluripotent stem cells. Here we establish a robust and efficient protocol for the rapid production of microglia-like cells from human (h) embryonic stem (ES) and induced pluripotent stem (iPS) cells that uses defined serum-free culture conditions...
September 26, 2016: Nature Medicine
Andreas Weltin, Steffen Hammer, Fozia Noor, Yeda Kaminski, Jochen Kieninger, Gerald A Urban
3D hepatic microtissues, unlike 2D cell cultures, retain many of the in-vivo-like functionalities even after long-term cultivation. Such 3D cultures are increasingly applied to investigate liver damage due to drug exposure in toxicology. However, there is a need for thorough metabolic characterization of these microtissues for mechanistic understanding of effects on culture behaviour. We measured metabolic parameters from single human HepaRG hepatocyte spheroids online and continuously with electrochemical microsensors...
July 28, 2016: Biosensors & Bioelectronics
Sonja Kallendrusch, Nicolas Schopow, Sonja C Stadler, Hildegard Büning, Ulrich Hacker
Adipose tissue plays a pivotal role, both in the regulation of energy homeostasis and as an endocrine organ. Consequently, adipose tissue dysfunction is closely related to insulin resistance, morbid obesity and the metabolic syndrome. To study molecular mechanisms and to develop novel therapeutic strategies, techniques are required to genetically modify mature adipocytes. Here, we report on adeno-associated viral vectors (rAAV) as a versatile tool to transduce human mature adipocytes in organotypic 3-dimensional (3-D) tissue cultures...
September 21, 2016: Human Gene Therapy Methods
Gili Kaufman, Drago Skrtic
Cariopathogen Streptococcus mutans (S. mutans) exists in infected dental pulp of deciduous teeth and is frequently linked with heart diseases. Organotypic (3D) dental pulp stem cells (DPSCs) cultures/microtissues, developed to mimic the physiological conditions in vivo, were utilized to assess the bacterial impact on their (i) 3D structural configuration, and (ii) recovery mechanisms. The cultures, developed in extracellular matrix (ECM) bio-scaffold (MatrigelTM), interacted with Wild type and GFP-tagged bacterial biofilms by permitting their infiltration through the ECM...
September 14, 2016: Journal of Medical Microbiology
A Tapia-Bustos, R Perez-Lobos, V Vío, C Lespay-Rebolledo, E Palacios, A Chiti-Morales, D Bustamante, M Herrera-Marschitz, P Morales
Perinatal asphyxia (PA) is associated to delayed cell death, affecting neurocircuitries of basal ganglia and hippocampus, and long-term neuropsychiatric disabilities. Several compensatory mechanisms have been suggested to take place, including cell proliferation and neurogenesis. There is evidence that PA can increase postnatal neurogenesis in hippocampus and subventricular zone (SVZ), modulated by dopamine, by still unclear mechanisms. We have studied here the effect of selective dopamine receptor agonists on cell death, cell proliferation and neurogenesis in organotypic cultures from control and asphyxia-exposed rats...
September 16, 2016: Neurotoxicity Research
Erin F Spence, Daniel J Kanak, Benjamin R Carlson, Scott H Soderling
UNLABELLED: Dendritic filopodia are actin-rich structures that are thought to contribute to early spine synapse formation; however, the actin regulatory proteins important for early synaptogenesis are poorly defined. Using organotypic hippocampal slice cultures and primary neuron hippocampal cultures from Arp2/3 conditional knock-out mice, we analyze the roles of the Arp2/3 complex, an actin regulator that creates branched actin networks, and demonstrate it is essential for distinct stages of both structural and functional maturation of excitatory spine synapses...
September 14, 2016: Journal of Neuroscience: the Official Journal of the Society for Neuroscience
Andrew P Patton, Johanna E Chesham, Michael H Hastings
UNLABELLED: The suprachiasmatic nucleus (SCN) is the master circadian oscillator encoding time-of-day information. SCN timekeeping is sustained by a cell-autonomous transcriptional-translational feedback loop, whereby expression of the Period and Cryptochrome genes is negatively regulated by their protein products. This loop in turn drives circadian oscillations in gene expression that direct SCN electrical activity and thence behavior. The robustness of SCN timekeeping is further enhanced by interneuronal, circuit-level coupling...
September 7, 2016: Journal of Neuroscience: the Official Journal of the Society for Neuroscience
Rachel J Watkins, Waraporn Imruetaicharoenchoke, Martin L Read, Neil Sharma, Vikki L Poole, Erica Gentillin, Sukhchain Bansal, Emy Bosseboeuf, Rachel Fletcher, Hannah R Nieto, Ujjal Mallick, Allan Hackshaw, Hisham Mehanna, Kristien Boelaert, Vicki E Smith, Christopher J McCabe
CONTEXT: Metastatic disease is responsible for the majority of endocrine cancer deaths. New therapeutic targets are urgently needed to improve patient survival rates. OBJECTIVE: The proto-oncogene PTTG1-Binding Factor (PBF/PTTG1IP) is overexpressed in multiple endocrine cancers and circumstantially associated with tumour aggressiveness. This study aimed to understand the role of PBF in tumour cell invasion and identify possible routes to inhibit its action. Design, Setting, Patients, and Interventions: Thyroid, breast and colorectal cells were transfected with PBF and cultured for in vitro analysis...
September 7, 2016: Journal of Clinical Endocrinology and Metabolism
Zeinab Ghanavati, Mahmoud Orazizadeh, Vahid Bayati, Mohammad Reza Abbaspour, Layasadat Khorsandi, Esrafil Mansouri, Niloofar Neisi
OBJECTIVE: The organotypic co-culture is a well-known technique to examine cellular interactions and their roles in stem cell proliferation and differentiation. This study aims to evaluate the effects of dermal fibroblasts (DFs) on epidermal differentiation of adipose-derived stem cells (ASCs) using a three-dimensional (3D) organotypic co- culture technique. MATERIALS AND METHODS: In this experimental research study, rat DFs and ASCs were isolated and cultured separately on electrospun polycaprolactone (PCL) matrices...
2016: Cell Journal
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