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Mammalian cell culture

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https://www.readbyqxmd.com/read/29352171/a-urine-dependent-human-urothelial-organoid-offers-a-potential-alternative-to-rodent-models-of-infection
#1
Harry Horsley, Dhanuson Dharmasena, James Malone-Lee, Jennifer L Rohn
Murine models describe a defined host/pathogen interaction for urinary tract infection, but human cell studies are scant. Although recent human urothelial organoid models are promising, none demonstrate long-term tolerance to urine, the natural substrate of the tissue and of the uropathogens that live there. We developed a novel human organoid from progenitor cells which demonstrates key structural hallmarks and biomarkers of the urothelium. After three weeks of transwell culture with 100% urine at the apical interface, the organoid stratified into multiple layers...
January 19, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29352169/chaski-a-novel-drosophila-lactate-pyruvate-transporter-required-in-glia-cells-for-survival-under-nutritional-stress
#2
María Graciela Delgado, Carlos Oliva, Estefanía López, Andrés Ibacache, Alex Galaz, Ricardo Delgado, L Felipe Barros, Jimena Sierralta
The intercellular transport of lactate is crucial for the astrocyte-to-neuron lactate shuttle (ANLS), a model of brain energetics according to which neurons are fueled by astrocytic lactate. In this study we show that the Drosophila chaski gene encodes a monocarboxylate transporter protein (MCT/SLC16A) which functions as a lactate/pyruvate transporter, as demonstrated by heterologous expression in mammalian cell culture using a genetically encoded FRET nanosensor. chaski expression is prominent in the Drosophila central nervous system and it is particularly enriched in glia over neurons...
January 19, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29348392/rapamycin-prevents-the-intervertebral-disc-degeneration-via-inhibiting-differentiation-and-senescence-of-annulus-fibrosus-cells
#3
Changhong Gao, Bin Ning, Chenglin Sang, Ying Zhang
The effects of bleomycin and rapamycin on cellular senescence and differentiation of rabbit annulus fibrosus stem cells (AFSCs) were investigated using a cell culture model. The results showed that bleomycin induced cellular senescence in AFSCs as evidenced by senescence-associated secretory phenotype. The morphology of AFSCs was changed from cobblestone-like cells to pancake-like cells. The senescence-associated β-galactosidase activity, the protein expression of P16 and P21, and inflammatory-related marker gene levels IL-1β, IL-6, and TNF-α were increased in bleomycin-treated AFSCs in a dose-dependent manner...
January 18, 2018: Aging
https://www.readbyqxmd.com/read/29347346/elucidating-distinct-ion-channel-populations-on-the-surface-of-hippocampal-neurons-via-single-particle-tracking-recurrence-analysis
#4
Grzegorz Sikora, Agnieszka Wyłomańska, Janusz Gajda, Laura Solé, Elizabeth J Akin, Michael M Tamkun, Diego Krapf
Protein and lipid nanodomains are prevalent on the surface of mammalian cells. In particular, it has been recently recognized that ion channels assemble into surface nanoclusters in the soma of cultured neurons. However, the interactions of these molecules with surface nanodomains display a considerable degree of heterogeneity. Here, we investigate this heterogeneity and develop statistical tools based on the recurrence of individual trajectories to identify subpopulations within ion channels in the neuronal surface...
December 2017: Physical Review. E
https://www.readbyqxmd.com/read/29346283/the-biological-role-of-hyaluronan-rich-oocyte-cumulus-extracellular-matrix-in-female-reproduction
#5
REVIEW
Eva Nagyova
Fertilization of the mammalian oocyte requires interactions between spermatozoa and expanded cumulus extracellular matrix (ECM) that surrounds the oocyte. This review focuses on key molecules that play an important role in the formation of the cumulus ECM, generated by the oocyte-cumulus complex. In particular, the specific inhibitors (AG1478, lapatinib, indomethacin and MG132) and progesterone receptor antagonist (RU486) exerting their effects through the remodeling of the ECM of the cumulus cells surrounding the oocyte have been described...
January 18, 2018: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/29344656/liraglutide-suppresses-proliferation-and-induces-adipogenic-differentiation-of-3t3-l1-cells-via-the-hippo-yap-signaling-pathway
#6
Yongmei Li, Jianying Du, Endong Zhu, Juanjuan Zhang, Jie Han, Wei Zhao, Bei Sun, Derun Tian
Liraglutide, as a glucagon-like peptide‑1 analogue, is used to treat type 2 diabetes mellitus and obesity. Previous findings have demonstrated the effects of liraglutide on adipogenesis; however, the underlying mechanism involved in this process remains to be elucidated. In the present study, to certify the effect of liraglutide on adipogenesis and explore the possible underlying mechanism involved in this process, preadipocyte 3T3‑L1 cells were cultured in adipocyte‑inducing medium and treated with liraglutide...
January 16, 2018: Molecular Medicine Reports
https://www.readbyqxmd.com/read/29342179/the-polymethoxy-flavonoid-sudachitin-suppresses-inflammatory-bone-destruction-by-directly-inhibiting-osteoclastogenesis-due-to-reduced-ros-production-and-mapk-activation-in-osteoclast-precursors
#7
Yoko Ohyama, Junta Ito, Victor J Kitano, Jun Shimada, Yoshiyuki Hakeda
Inflammatory bone diseases, including rheumatoid arthritis, periodontitis and peri-implantitis, are associated not only with the production of inflammatory cytokines but also with local oxidative status, which is defined by intracellular reactive oxygen species (ROS). Osteoclast differentiation has been reported to be related to increased intracellular ROS levels in osteoclast lineage cells. Sudachitin, which is a polymethoxyflavone derived from Citrus sudachi, possesses antioxidant properties and regulates various functions in mammalian cells...
2018: PloS One
https://www.readbyqxmd.com/read/29336068/isatin-an-endogenous-nonpeptide-biofactor-a-review-of-its-molecular-targets-mechanisms-of-actions-and-their-biomedical-implications
#8
REVIEW
Alexei Medvedev, Olga Buneeva, Oksana Gnedenko, Pavel Ershov, Alexis Ivanov
Isatin (indole-2,3-dione) is an oxidized indole. It is widely distributed in mammalian tissues and body fluids, where isatin concentrations vary significantly from <0.1 to > 10 µM. Isatin output is increased under conditions of stress. Exogenously administered isatin is characterized by low toxicity, mutagenicity, and genotoxicity in vivo. Cytotoxic effects of isatin on various cell cultures are usually observed at concentrations exceeding 100 µM. Binding of [3 H]isatin to rat brain sections is consistent with its physiological concentrations...
January 16, 2018: BioFactors
https://www.readbyqxmd.com/read/29335484/microbial-competition-of-rhodotorula-mucilaginosa-uanl-001l-and-e-coli-increase-biosynthesis-of-non-toxic-exopolysaccharide-with-applications-as-a-wide-spectrum-antimicrobial
#9
Augusto Vazquez-Rodriguez, Ximena G Vasto-Anzaldo, Daniel Barboza Perez, Eduardo Vázquez-Garza, Héctor Chapoy-Villanueva, Gerardo García-Rivas, Javier A Garza-Cervantes, Jéssica J Gómez-Lugo, Alma Elizabeth Gomez-Loredo, Maria Teresa Garza Gonzalez, Xristo Zarate, Jose Ruben Morones-Ramirez
Bacterial species are able to colonize and establish communities in biotic and abiotic surfaces. Moreover, within the past five decades, incidence of bacterial strains resistant to currently used antibiotics has increased dramatically. This has led to diverse health issues and economical losses for different industries. Therefore, there is a latent need to develop new and more efficient antimicrobials. This work reports an increased production of an exopolysaccharide in a native yeast strain isolated from the Mexican Northeast, Rhodotorula mucilaginosa UANL-001L, when co-cultured with E...
January 15, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29334988/epidermal-wnt-signalling-regulates-transcriptome-heterogeneity-and-proliferative-fate-in-neighbouring-cells
#10
Arsham Ghahramani, Giacomo Donati, Nicholas M Luscombe, Fiona M Watt
BACKGROUND: Canonical Wnt/beta-catenin signalling regulates self-renewal and lineage selection within the mammalian epidermis. Although the transcriptional response of keratinocytes that receive a Wnt signal is well characterized, little is known about the mechanism by which keratinocytes in proximity to the Wnt-receiving cell are co-opted to undergo a change in cell fate. RESULTS: To address this, we perform single-cell RNA-sequencing on mouse keratinocytes co-cultured with and without beta-catenin-activated neighbouring cells...
January 15, 2018: Genome Biology
https://www.readbyqxmd.com/read/29332592/construction-of-an-m1gs-ribozyme-for-targeted-and-rapid-mrna-cleavage-application-on-the-ets-2-oncogene
#11
Chrisavgi Toumpeki, Dimitrios Anastasakis, Ioannis Panagoulias, Vassiliki Stamatopoulou, Tassos Georgakopoulos, Sofia Kallia-Raftopoulos, Athanasia Mouzaki, Denis Drainas
Background RNase P-mediated cleavage of target RNAs has been proposed as a promising tool for gene silencing. Ets-2 proto-oncogene controls the expression of a wide variety of genes involved in cancer and immunity. Objective Construction of a functional RNase P-based ribozyme (M1GS303) that targets Ets-2 mRNA Method The accessible sites for targeting of Ets-2 mRNA were identified by footprinting analysis. M1GS303 ribozyme was constructed by cloning. The activity of the ribozyme in the presence or absence of spiramysin in E...
January 11, 2018: Medicinal Chemistry
https://www.readbyqxmd.com/read/29331783/comparative-study-of-novel-in-situ-decorated-porous-chitosan-selenium-scaffolds-and-porous-chitosan-silver-scaffolds-towards-antimicrobial-wound-dressing-application
#12
Dhee P Biswas, Neil M O'Brien-Simpson, Eric C Reynolds, Andrea J O'Connor, Phong A Tran
Dermal defects caused by trauma or disease are challenging to treat due to difficult-to-treat infections that impair wound healing. Due to the widespread emergence of drug-resistant bacteria and dwindling discoveries of new antibiotics, there is currently an urgent need to introduce novel antimicrobials effective against antibiotic-resistant bacteria without causing damage to host tissues. As selenium (Se) and silver (Ag) are known for their antimicrobial properties, we investigated the separate loading of these materials into porous chitosan/PVA (CS) scaffolds through a simple in situ deposition method to create two distinct wound dressing materials (CS-Se and CS-Ag)...
January 5, 2018: Journal of Colloid and Interface Science
https://www.readbyqxmd.com/read/29328421/antitumor-activity-of-curcumin-by-modulation-of-apoptosis-and-autophagy-in-human-lung-cancer-a549-cells-through-inhibiting-pi3k-akt-mtor-pathway
#13
Furong Liu, Song Gao, Yuxuan Yang, Xiaodan Zhao, Yameng Fan, Wenxia Ma, Danrong Yang, Aimin Yang, Yan Yu
Curcumin is known to exhibit anticancer effects on various cancers with selective cytotoxicity in tumor cells. In the present study, the effects of curcumin‑induced multiple PCDs on human non‑small cell lung cancer (NSCLC) cells and the potential molecular mechanisms of apoptosis and autophagy triggered by curcumin via the PI3K/Akt/mTOR signaling pathway were explored, further confirmed by co‑culture of curcumin with mTOR blocker rapamycin and PI3K/Akt inhibitor LY294002. The anti‑proliferation effect of different stimulus was measured by MTT assay...
January 4, 2018: Oncology Reports
https://www.readbyqxmd.com/read/29325543/persistent-mammalian-orthoreovirus-coxsackievirus-and-adenovirus-co-infection-in-a-child-with-a-primary-immunodeficiency-detected-by-metagenomic-sequencing-a-case-report
#14
Dagmara W Lewandowska, Riccarda Capaul, Seraina Prader, Osvaldo Zagordi, Fabienne-Desirée Geissberger, Martin Kügler, Marcus Knorr, Christoph Berger, Tayfun Güngör, Janine Reichenbach, Cyril Shah, Jürg Böni, Andrea Zbinden, Alexandra Trkola, Jana Pachlopnik Schmid, Michael Huber
BACKGROUND: We report a rare case of Mammalian orthoreovirus (MRV) infection in a child with a primary immunodeficiency (PID). Infections with Mammalian orthoreovirus are very rare and probably of zoonotic origin. Only a few cases have been described so far, including one with similar pathogenesis as in our case. CASE PRESENTATION: The patient, age 11, presented with flu-like symptoms and persistent severe diarrhea. Enterovirus has been detected over several months, however, exact typing of a positive cell culture remained inconclusive...
January 11, 2018: BMC Infectious Diseases
https://www.readbyqxmd.com/read/29321321/functional-analysis-of-the-dengue-virus-genome-using-an-insertional-mutagenesis-screen
#15
Jeffrey W Perry, Yanhua Chen, Elizabeth Speliotes, Andrew W Tai
In the last few decades, dengue virus, an arbovirus, has spread to over 120 countries. Although a vaccine has been approved in some countries, limitations on its effectiveness, and a lack of effective antiviral treatments reinforce the need for additional research. The functions of several viral nonstructural proteins are essentially unknown. To better understand the functions of these proteins and thus dengue virus pathogenesis, we embarked on a genome wide transposon mutagenesis screen with Next Generation Sequencing to determine sites in the viral genome that tolerate 15 nucleotide insertions...
January 10, 2018: Journal of Virology
https://www.readbyqxmd.com/read/29318876/on-demand-modulation-of-bacterial-cell-fates-on-multifunctional-dynamic-substrates
#16
Inseong Choi, Jinhwan Lee, Wontae Kim, Hyunook Kang, Se Won Bae, Rakwoo Chang, Sunghyun Kim, Woon-Seok Yeo
This paper reports unprecedented dynamic surfaces based on zwitterionic low density self-assembled monolayers (LDSAMs) of alkanethiolates on gold, which integrate three interconvertible states - bacteria-adherable, bactericidal, and nonfouling states - through electrical modulations. The conformations of alkanethiolates were electrically modulated to generate zwitterionic, anionic, and cationic surfaces, which responded differently to bacteria and determined the fate of bacteria. Furthermore, the reversible switching of multi-functions of the surface was realized for killing bacteria and subsequently releasing dead bacteria from the surface...
January 10, 2018: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/29314777/concomitant-reduction-of-lactate-and-ammonia-accumulation-in-fed-batch-cultures-impact-on-glycoprotein-production-and-quality
#17
Eric Karengera, Anna Robotham, John Kelly, Yves Durocher, Gregory De Crescenzo, Olivier Henry
Lactate and ammonia accumulation is a major factor limiting the performance of fed-batch strategies for mammalian cell culture processes. In addition to the detrimental effects of these by-products on production yield, ammonia also contributes to recombinant glycoprotein quality deterioration. In this study, we tackled the accumulation of these two inhibiting metabolic wastes by culturing in glutamine-free fed-batch cultures an engineered HEK293 cell line displaying an improved central carbon metabolism. Batch cultures highlighted the ability of PYC2-overexpressing HEK293 cells to grow and sustain a relatively high viability in absence of glutamine without prior adaptation to the culture medium...
January 5, 2018: Biotechnology Progress
https://www.readbyqxmd.com/read/29313356/sub-cellular-nanorheology-reveals-lysosomal-viscosity-as-a-reporter-for-lysosomal-storage-diseases
#18
John Devany, Kasturi Chakraborty, Yamuna Krishnan
We describe a new method to measure viscosity within sub-cellular organelles of a living cell using nanorheology. We demonstrate proof of concept by measuring viscosity in lysosomes in multiple cell types and disease models. The lysosome is an organelle, which is responsible for the breakdown of complex biomolecules. When different lysosomal proteins are defective, they are unable to breakdown specific biological substrates, that get stored within the lysosome, causing about 70 fatal diseases called lysosomal storage disorders (LSDs)...
January 9, 2018: Nano Letters
https://www.readbyqxmd.com/read/29313024/evolving-trends-in-mab-production-processes
#19
REVIEW
Abhinav A Shukla, Leslie S Wolfe, Sigma S Mostafa, Carnley Norman
Monoclonal antibodies (mAbs) have established themselves as the leading biopharmaceutical therapeutic modality. The establishment of robust manufacturing platforms are key for antibody drug discovery efforts to seamlessly translate into clinical and commercial successes. Several drivers are influencing the design of mAb manufacturing processes. The advent of biosimilars is driving a desire to achieve lower cost of goods and globalize biologics manufacturing. High titers are now routinely achieved for mAbs in mammalian cell culture...
March 2017: Bioengineering & Translational Medicine
https://www.readbyqxmd.com/read/29305410/evolution-of-tools-and-methods-for-monitoring-autophagic-flux-in-mammalian-cells
#20
REVIEW
Kevin C Yang, Paalini Sathiyaseelan, Cally Ho, Sharon M Gorski
Autophagy is an evolutionarily conserved lysosome-mediated degradation and recycling process, which functions in cellular homeostasis and stress adaptation. The process is highly dynamic and involves autophagosome synthesis, cargo recognition and transport, autophagosome-lysosome fusion, and cargo degradation. The multistep nature of autophagy makes it challenging to quantify, and it is important to consider not only the number of autophagosomes within a cell but also the autophagic degradative activity. The rate at which cargos are recognized, segregated, and degraded through the autophagy pathway is defined as autophagic flux...
January 5, 2018: Biochemical Society Transactions
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