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https://www.readbyqxmd.com/read/27922179/a-carbon-dioxide-stripping-model-for-mammalian-cell-culture-in-manufacturing-scale-bioreactors
#1
Zizhuo Xing, Amanda M Lewis, Michael C Borys, Zheng Jian Li
Control of carbon dioxide within the optimum range is important in mammalian bioprocesses at the manufacturing scale in order to ensure robust cell growth, high protein yields, and consistent quality attributes. The majority of bioprocess development work is done in laboratory bioreactors, in which carbon dioxide levels are more easily controlled. Some challenges in carbon dioxide control can present themselves when cell culture processes are scaled up, because carbon dioxide accumulation is a common feature due to longer gas-residence time of mammalian cell culture in large scale bioreactors...
December 6, 2016: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/27915978/cell-compatibility-of-an-eposimal-vector-mediated-by-the-characteristic-motifs-of-matrix-attachment-regions
#2
Tian-Yun Wang, Li Wang, Yu-Xin Yang, Chun-Peng Zhao, Yan-Long Jia, Qin Li, Jun-He Zhang, Yi-You Peng, Miao Wang, Hong-Yan Xu, Xiao-Yin Wang
The characteristic sequence of β-interferon matrix attachment regions (MARs) can mediate transgene expression via episomal vectors in Chinese hamster ovary (CHO) cells. However, the host cells were from hamster ovaries, which are not suitable target cells for gene therapy. In this study, we aimed to evaluate the suitability of 12 different human cell lines as target cells for gene therapy. We transfected the cells with episomal vectors and obtained colonies stably expressing the vector products after G418 screening...
December 2, 2016: Current Gene Therapy
https://www.readbyqxmd.com/read/27909054/identification-of-a-substrate-selective-exosite-within-the-metalloproteinase-anthrax-lethal-factor
#3
Allison B Goldberg, Eunice Cho, Chad J Miller, Hua Jane Lou, Benjamin E Turk
The metalloproteinase anthrax lethal factor (LF) is secreted by Bacillus anthracis to promote disease virulence through disruption of host signaling pathways. LF is a highly specific protease, exclusively cleaving mitogen-activated protein kinase kinases (MKKs) and rodent NACHT leucine-rich repeat and pyrin domain containing protein 1B (NLRP1B). How LF achieves such restricted substrate specificity is not understood. Previous studies have suggested the existence of an exosite interaction between LF and MKKs that promotes cleavage efficiency and specificity...
December 1, 2016: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/27907214/derivation-of-induced-trophoblast-cell-lines-in-cattle-by-doxycycline-inducible-piggybac-vectors
#4
Takamasa Kawaguchi, Dooseon Cho, Masafumi Hayashi, Tomoyuki Tsukiyama, Koji Kimura, Shuichi Matsuyama, Naojiro Minami, Masayasu Yamada, Hiroshi Imai
Trophectoderm lineage specification is one of the earliest differentiation events in mammalian development. The trophoblast lineage, which is derived from the trophectoderm, mediates implantation and placental formation. However, the processes involved in trophoblastic differentiation and placental formation in cattle remain unclear due to interspecies differences when compared with other model systems and the small repertoire of available trophoblast cell lines. Here, we describe the generation of trophoblast cell lines (biTBCs) from bovine amnion-derived cells (bADCs) using an induced pluripotent stem cell technique...
2016: PloS One
https://www.readbyqxmd.com/read/27906496/aerobic-expression-of-vitreoscilla-hemoglobin-improves-the-growth-performance-of-cho-k1-cells
#5
Mariana Juárez, Claudia H González-De la Rosa, Elisa Memún, Juan-Carlos Sigala, Alvaro R Lara
Inefficient carbon metabolism is a relevant issue during the culture of mammalian cells for the production of biopharmaceuticals. Therefore, cell engineering strategies to improve the metabolic and growth performance of cell lines are needed. The expression of Vitreoscilla stercoraria hemoglobin (VHb) has been shown to significantly reduce overflow metabolism and improve the aerobic growth of bacteria. However, the effects of VHb on mammalian cells have been rarely studied. Here, the impact of VHb on growth and lactate accumulation during CHO-K1 cell culture was investigated...
December 1, 2016: Biotechnology Journal
https://www.readbyqxmd.com/read/27900626/selection-of-chemically-defined-media-for-cho-cell-fed-batch-culture-processes
#6
Xiao Pan, Mathieu Streefland, Ciska Dalm, René H Wijffels, Dirk E Martens
Two CHO cell clones derived from the same parental CHO(BC®) cell line and producing the same monoclonal antibody (BC-G, a low producing clone; BC-P, a high producing clone) were tested in four basal media in all possible combinations with three feeds (=12 conditions) in fed-batch cultures. Higher amino acid feeding did not always lead to higher mAb production. The two clones showed differences in cell physiology, metabolism and optimal medium-feed combinations. During the phase transitions of all cultures, cell metabolism showed a shift represented by lower specific consumption and production rates, except for the specific glucose consumption rate in cultures fed by Actifeed A/B...
November 29, 2016: Cytotechnology
https://www.readbyqxmd.com/read/27897167/ecm-proteins-in-a-microporous-scaffold-influence-hepatocyte-morphology-function-and-gene-expression
#7
Yan Wang, Myung Hee Kim, Hitomi Shirahama, Jae Ho Lee, Soon Seng Ng, Jeffrey S Glenn, Nam-Joon Cho
It is well known that a three-dimensional (3D) culture environment and the presence of extracellular matrix (ECM) proteins facilitate hepatocyte viability and maintenance of the liver-specific phenotype in vitro. However, it is not clear whether specific ECM components such as collagen or fibronectin differentially regulate such processes, especially in 3D scaffolds. In this study, a series of ECM-functionalized inverted colloidal crystal (ICC) microporous scaffolds were fabricated and their influence on Huh-7...
November 29, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27877076/cardiomyogenic-differentiation-of-human-dental-follicle-derived-stem-cells-by-suberoylanilide-hydroxamic-acid-and-their-in-vivo-homing-property
#8
Iel-Yong Sung, Han-Na Son, Imran Ullah, Dinesh Bharti, Ju-Mi Park, Yeong-Cheol Cho, June-Ho Byun, Young-Hoon Kang, Su-Jin Sung, Jong-Woo Kim, Gyu-Jin Rho, Bong-Wook Park
The purpose of the present study was to investigate the in vitro cardiomyogenic differentiation potential of human dental follicle-derived stem cells (DFCs) under the influence of suberoylanilide hydroxamic acid (SAHA), a member of the histone deacetylase inhibitor family, and analyze the in vivo homing capacity of induced cardiomyocytes (iCMs) when transplanted systemically. DFCs from extracted wisdom teeth showed mesenchymal stem cell (MSC) characteristics such as plate adherent growing, expression of MSC markers (CD44, CD90, and CD105), and mesenchymal lineage-specific differentiation potential...
2016: International Journal of Medical Sciences
https://www.readbyqxmd.com/read/27877072/cultured-human-periosteum-derived-cells-can-differentiate-into-osteoblasts-in-a-perioxisome-proliferator-activated-receptor-gamma-mediated-fashion-via-bone-morphogenetic-protein-signaling
#9
Jin-Eun Chung, Jin-Ho Park, Jeong-Won Yun, Young-Hoon Kang, Bong-Wook Park, Sun-Chul Hwang, Yeong-Cheol Cho, Iel-Yong Sung, Dong Kyun Woo, June-Ho Byun
The differentiation of mesenchymal stem cells towards an osteoblastic fate depends on numerous signaling pathways, including activation of bone morphogenetic protein (BMP) signaling components. Commitment to osteogenesis is associated with activation of osteoblast-related signal transduction, whereas inactivation of this signal transduction favors adipogenesis. BMP signaling also has a critical role in the processes by which mesenchymal stem cells undergo commitment to the adipocyte lineage. In our previous study, we demonstrated that an agonist of the perioxisome proliferator-activated receptor γ (PPARγ), a master regulator of adipocyte differentiation, stimulates osteoblastic differentiation of cultured human periosteum-derived cells...
2016: International Journal of Medical Sciences
https://www.readbyqxmd.com/read/27869744/protective-effect-of-tempol-against-cisplatin-induced-ototoxicity
#10
Cha Kyung Youn, Jun Kim, Eu-Ri Jo, Jeonghyun Oh, Nam Yong Do, Sung Il Cho
One of the major adverse effects of cisplatin chemotherapy is hearing loss. Cisplatin-induced ototoxicity hampers treatment because it often necessitates dose reduction, which decreases cisplatin efficacy. This study was performed to investigate the effect of Tempol on cisplatin-induced ototoxicity in an auditory cell line, House Ear Institute-Organ of Corti 1 (HEI-OC1). Cultured HEI-OC1 cells were exposed to 30 μM cisplatin for 24 h with or without a 2 h pre-treatment with Tempol. Cell viability was determined using 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay and apoptotic cells were identified using terminal deoxynucleotidyl transferase dUTP nick end labeling of nuclei (TUNEL) assay and flow cytometry...
November 18, 2016: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/27869296/segmented-linear-modeling-of-cho-fed-batch-culture-and-its-application-to-large-scale-production
#11
Bassem Ben Yahia, Boris Gourevitch, Laetitia Malphettes, Elmar Heinzle
We describe a systematic approach to model CHO metabolism during biopharmaceutical production across a wide range of cell culture conditions. To this end, we applied the metabolic steady state concept. We analyzed and modeled the production rates of metabolites as a function of the specific growth rate. First, the total number of metabolic steady state phases and the location of the breakpoints were determined by recursive partitioning. For this, the smoothed derivative of the metabolic rates with respect to the growth rate were used followed by hierarchical clustering of the obtained partition...
November 7, 2016: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/27869292/model-based-investigation-of-intracellular-processes-determining-antibody-fc-glycosylation-under-mild-hypothermia
#12
Si Nga Sou, Philip M Jedrzejewski, Ken Lee, Christopher Sellick, Karen M Polizzi, Cleo Kontoravdi
Despite the positive effects of mild hypothermic conditions on monoclonal antibody (mAb) productivity (qmAb ) during mammalian cell culture, the impact of reduced culture temperature on mAb Fc-glycosylation and the mechanism behind changes in the glycan composition is not fully established. The lack of knowledge about the regulation of dynamic intracellular processes under mild hypothermia restricts bioprocess optimisation. To address this issue, a mathematical model that quantitatively describes CHO cell behaviour and metabolism, mAb synthesis and its N-linked glycosylation profiles before and after the induction of mild hypothermia is constructed using two sets of parameters...
November 21, 2016: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/27860372/biomaterials-and-culture-technologies-for-regenerative-therapy-of-liver-tissue
#13
Roman A Perez, Cho-Rok Jung, Hae-Won Kim
Regenerative approach has emerged to substitute the current extracorporeal technologies for the treatment of diseased and damaged liver tissue. This is based on the use of biomaterials that modulate the responses of hepatic cells through the unique matrix properties tuned to recapitulate regenerative functions. Cells in liver preserve their phenotype or differentiate through the interactions with extracellular matrix molecules. Therefore, the intrinsic properties of the engineered biomaterials, such as stiffness and surface topography, need to be tailored to induce appropriate cellular functions...
November 18, 2016: Advanced Healthcare Materials
https://www.readbyqxmd.com/read/27853628/tir-domain-containing-adapter-inducing-interferon-%C3%AE-trif-regulates-cxcr5-t-helper-cells-in-the-intestine
#14
Saravana Kanagavelu, Claudia Flores, Shinichiro Hagiwara, Jose Ruiz, Jinhee Hyun, Ei E Cho, Frank Sun, Laura Romero, David Q Shih, Masayuki Fukata
OBJECTIVE: Establishing an effective humoral immunity is an important host defense mechanism in intestinal mucosa. T follicular helper (Tfh) cells are a spectrum of CXCR5 expressing T helper cells that induce antigen-specific B cell differentiation. Because the differentiation of T helper cells is largely regulated by innate immunity, we addressed whether TRIF signaling regulates Tfh cell differentiation and its ability to trigger humoral immune responses in the intestine. METHOD: CD4(+)CXCR5(+) T cells, B cells, and plasma cells in the Peyer's patches (PPs) of WT and TRIF-deficient (Trif(LPS2)) mice were analyzed by flow cytometry at the baseline, 9 days post primary infection, and 7 days post-secondary infection with Y...
October 2016: Journal of Clinical & Cellular Immunology
https://www.readbyqxmd.com/read/27847750/in-vitro-characterization-of-human-dental-pulp-stem-cells-isolated-by-three-different-methods
#15
Ji-Hyun Jang, Hyeon-Woo Lee, Kyu Min Cho, Hee-Woong Shin, Mo Kwan Kang, Sang Hyuk Park, Euiseong Kim
OBJECTIVES: In this study, we characterized human dental pulp cells (HDPCs) obtained by different culture methods to establish the most suitable methodology for dental tissue engineering and regenerative endodontic applications. MATERIALS AND METHODS: HDPCs were isolated by the outgrowth method (HDPCs-OG), the enzymatic digestion method (collagenase/dispase/trypsin, HDPCs-ED), or the combination of both methods (HDPCs-Combined). The expression of mesenchymal stem cell markers (CD105, CD90, and CD73) was investigated...
November 2016: Restorative Dentistry & Endodontics
https://www.readbyqxmd.com/read/27832969/advances-in-analytical-methodologies-to-guide-bioprocess-engineering-for-bio-therapeutics
#16
Radka Saldova, Michelle Kilcoyne, Henning Stöckmann, Silvia Millán Martín, Amanda M Lewis, Catherine M E Tuite, Jared Q Gerlach, Marie Le Berre, Michael C Borys, Zheng Jian Li, Nicholas R Abu-Absi, Kirk Leister, Lokesh Joshi, Pauline M Rudd
This study was performed to monitor the glycoform distribution of a recombinant antibody fusion protein expressed in CHO cells over the course of fed-batch bioreactor runs using high-throughput methods to accurately determine the glycosylation status of the cell culture and its product. Three different bioreactors running similar conditions were analysed at the same six time-points using the advanced methods described here. N-glycans from cell and secreted glycoproteins from CHO cells were analysed by HILIC-UPLC and MS, and the total glycosylation (both N- and O-linked glycans) secreted from the CHO cells were analysed by lectin microarrays...
November 7, 2016: Methods: a Companion to Methods in Enzymology
https://www.readbyqxmd.com/read/27825930/novel-approach-to-study-the-cardiovascular-effects-and-mechanism-of-action-of-urban-particulate-matter-using-lung-epithelial-endothelial-tetra-culture-system
#17
Ha Ryong Kim, Han Soo Cho, Da Young Shin, Kyu Hyuck Chung
In vitro models have become increasingly sophisticated, and their usefulness in supporting toxicity testing is well established. The present study was designed to establish a novel in vitro model that mimics the cellular network surrounding airways and pulmonary blood vessels, to study the cardiovascular toxic effects of particulate matter (PM). Transwell culture method was used to develop a novel tetra-culture system consisting of tri-cultures (one lung epithelial and two immune cell lines) in the apical chamber and endothelial cells in the basolateral chamber...
November 4, 2016: Toxicology in Vitro: An International Journal Published in Association with BIBRA
https://www.readbyqxmd.com/read/27819051/serine-one-carbon-catabolism-with-formate-overflow
#18
Johannes Meiser, Sergey Tumanov, Oliver Maddocks, Christiaan Fred Labuschagne, Dimitris Athineos, Niels Van Den Broek, Gillian M Mackay, Eyal Gottlieb, Karen Blyth, Karen Vousden, Jurre J Kamphorst, Alexei Vazquez
Serine catabolism to glycine and a one-carbon unit has been linked to the anabolic requirements of proliferating mammalian cells. However, genome-scale modeling predicts a catabolic role with one-carbon release as formate. We experimentally prove that in cultured cancer cells and nontransformed fibroblasts, most of the serine-derived one-carbon units are released from cells as formate, and that formate release is dependent on mitochondrial reverse 10-CHO-THF synthetase activity. We also show that in cancer cells, formate release is coupled to mitochondrial complex I activity, whereas in nontransformed fibroblasts, it is partially insensitive to inhibition of complex I activity...
October 2016: Science Advances
https://www.readbyqxmd.com/read/27817046/production-and-purification-of-recombinant-glucocerebrosidase-in-transgenic-rice-cell-suspension-cultures
#19
Hyung-Jin Nam, Jun-Young Kwon, Hong-Yeol Choi, Seung-Hoon Kang, Hahn-Sun Jung, Dong-Il Kim
Gaucher disease, which is caused by deficiency of glucocerebrosidase (GCD), is currently treated by enzyme replacement therapy. Plant-based systems produce glycoproteins and can be combined with targeting strategies to generate proteins with terminal mannose structures for macrophage uptake. However, the gliding step for the purification is essential since the produced protein still exists inside cells. In the case of rice-amylase 1A (RAmy1A) secretion signal peptide, GCD protein is secreted outside of cells and simplifies the purification step...
November 5, 2016: Applied Biochemistry and Biotechnology
https://www.readbyqxmd.com/read/27811063/loss-of-function-screens-of-druggable-targetome-against-cancer-stem-like-cells
#20
Mee Song, Hani Lee, Myung-Hee Nam, Euna Jeong, Somin Kim, Yourae Hong, Nayoung Kim, Hwa Young Yim, Young-Ji Yoo, Jung Seok Kim, Jin-Seok Kim, Yong-Yeon Cho, Gordon B Mills, Woo-Young Kim, Sukjoon Yoon
Cancer stem-like cells (CSLCs) contribute to the initiation and recurrence of tumors and to their resistance to conventional therapies. In this study, small interfering RNA (siRNA)-based screening of ∼4800 druggable genes in 3-dimensional CSLC cultures in comparison to 2-dimensional bulk cultures of U87 glioma cells revealed 3 group of genes essential for the following: survival of the CSLC population only, bulk-cultured population only, or both populations. While diverse biologic processes were associated with siRNAs reducing the bulk-cultured population, CSLC-eliminating siRNAs were enriched in a few functional categories such as lipid metabolism, protein metabolism, and gene expression...
October 20, 2016: FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology
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