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CHO cell culture

Yao-Xiong Huang, Chun-Lan He, Ping Wang, Yan-Ting Pan, Wei-Wei Tuo, Cheng-Can Yao
We report here a versatile on-stage microfluidic cell culture and assay system which is compatible with different microscopes and sensors, can simultaneously perform steps of long term cell culture, high throughput time lapse cell assays/imaging, and cell micromanipulations. With the system, we cultured a variety of cells for different periods of time and monitored their cell morphology, migration and division. We also performed a series non-invasive real time in situ time lapse assays and micromanipulations on different cells...
October 4, 2017: Biosensors & Bioelectronics
Chih-Jung Hsu, Harsh V Jain, Abasha Williams, Julie Wang, Scott C Lute, Serge L Beaucage, Kurt A Brorson
A rapid and cost-effective transient transfection method for mammalian cells is essential for screening biopharmaceuticals in early stages of development. A library of 25 amphipathic trans-acting oligodeoxythymidine phosphorothioate triester (dTtaPS) transfection reagents, carrying positively charged and lipophilic groups, has been constructed for this purpose. High-throughput screening of the library, using an imaging cytometer and an automated microbioreactor system, has led to the identification of dTtaPS(10+) as a potent transfection reagent...
October 10, 2017: Biotechnology and Applied Biochemistry
Barbara Di Martino, Federica Di Profio, Irene Melegari, Vittorio Sarchese, Ivano Massirio, Alessia Luciani, Gianvito Lanave, Fulvio Marsilio, Vito Martella
Vesivirus 2117 was first discovered as a contaminant in Chinese hamster ovary (CHO) cell cultures used for human drug production. Similar vesiviruses (VeVs) have been detected recently in dogs. In order to address the hypothesis that cats may also be exposed to 2117-like VeVs, in this study, we screened 236 feline sera using an enzyme-linked immunosorbent assay (ELISA) based on a recombinant VP1 protein from the canine VeV Bari/212/07/ITA. IgG antibodies against the 2117-like VeV were detected in 37.3% of the sera tested...
October 11, 2017: Archives of Virology
Lilian Grigorian-Shamagian, Weixin Liu, Soraya Fereydooni, Ryan C Middleton, Jackelyn Valle, Jae Hyung Cho, Eduardo Marbán
Aim: The aim is to assess the effects of CDCs on heart structure, function, gene expression, and systemic parameters in aged rats. Diastolic dysfunction is characteristic of aged hearts. Cardiosphere-derived cell (CDC) therapy has exhibited several favourable effects on heart structure and function in humans and in preclinical models; however, the effects of CDCs on aging have not been evaluated. Methods and Results: We compared intra-cardiac injections of neonatal rat CDCs to vehicle (phosphate-buffered saline, PBS) in 21...
August 14, 2017: European Heart Journal
Seyedeh Hoda Jazayeri, Amir Amiri-Yekta, Hamid Gourabi, Baharak Abd Emami, Zahra Halfinezhad, Somayeh Abolghasemi, Nayeralsadat Fatemi, Abbas Daneshipour, Mohammad Hossein Ghahremani, Mohammad Hossein Sanati, Mohammad Reza Khorramizadeh
Production of recombinant pharmaceutical proteins has made a great contribution to modern biotechnology. At present, quick advances in protein expression lead to the enhancement of product quantity and quality as well as reduction in timescale processing. In the current study, we assessed the expression level of recombinant human follicle-stimulating hormone (rhFSH) in adherent and suspension Chinese hamster ovary (CHO) cell lines by cultivation in serum-containing and chemically defined, protein-free media...
October 9, 2017: Molecular Biotechnology
C Castillo, S Zaror, M Gonzalez, A Hidalgo, C F Burgos, O I Cabezas, F Hugues, S P Jiménez, E González-Horta, I González-Chavarría, J Gavilán, R Montesino, O Sánchez, Manuela G Lopez, J Fuentealba, J R Toledo
Human erythropoietin is mainly recognized for its hematopoietic function; however, by binding to its receptor (EpoR), it can activate different signaling pathways as STAT, PI3K, MAPK and RAS to increase cellular differentiation or provide neuroprotective effects, among others. A recombinant human erythropoietin variant with low glycosylation and without hematopoietic effect (EpoL) was purified from skimmed goat milk. Recombinant human erythropoietin (Epo) was obtained from CHO cell line and used as control to compare EpoL effects...
September 21, 2017: Redox Biology
Dang-Huan Pham, Ju-Sun Kim, Sang-Ki Kim, Dong-Jun Shin, Nguyen-Thanh-Tung Uong, Hoon Hyun, Mee Sun Yoon, Sin Jae Kang, Young Jae Ryu, Jin Seong Cho, Jung Han Yoon, Ji Shin Lee, Duck Cho, Soo-Hyeon Lee, Min Ho Park
BACKGROUND/AIM: The inhibition of a disintegrin and metalloproteinase (ADAM) has the potential to become a novel approach for natural killer (NK) cell-based cancer immunotherapy. Thus, the aim of this study was to investigate the influence of ADAM10 and ADAM17 inhibitors on expanded NK cell to enhance antibody-dependent cellular cytotoxicity (ADCC) in breast cancer cell lines. MATERIALS AND METHODS: NK cells were expanded in medium supplemented with an ADAM10 or ADAM17 inhibitor to prevent the shedding of soluble CD16/FcγRIII...
October 2017: Anticancer Research
Cynthia L Fisher, Hendrik Marks, Lily Ting-Yin Cho, Robert Andrews, Sam Wormald, Thomas Carroll, Vivek Iyer, Peri Tate, Barry Rosen, Hendrik G Stunnenberg, Amanda G Fisher, William C Skarnes
Mouse embryonic stem (ES) cells are a popular model system to study biological processes, though uncovering recessive phenotypes requires inactivating both alleles. Building upon resources from the International Knockout Mouse Consortium (IKMC), we developed a targeting vector for second allele inactivation in conditional-ready IKMC 'knockout-first' ES cell lines. We applied our technology to several epigenetic regulators, recovering bi-allelic targeted clones with a high efficiency of 60% and used Flp recombinase to restore expression in two null cell lines to demonstrate how our system confirms causality through mutant phenotype reversion...
September 13, 2017: Nucleic Acids Research
Ye-Seul Yoon, Eun-Duk Cho, Woo Jung Ahn, Kyung Won Lee, Seung-Jae Lee, He-Jin Lee
Autophagy is a pivotal intracellular process by which cellular macromolecules are degraded upon various stimuli. A failure in the degradation of autophagic substrates such as impaired organelles and protein aggregates leads to their accumulations, which are characteristics of many neurodegenerative diseases. Pharmacological activation of autophagy has thus been considered a prospective therapeutic approach for treating neurodegenerative diseases. Among a number of autophagy-inducing agents, trehalose has received attention for its beneficial effects in different disease models of neurodegeneration...
October 5, 2017: Cell Death & Disease
Alan Griffith, Paul S Kelly, Sebastian Vencken, Nga T Lao, Catherine M Greene, Martin Clynes, Niall Barron
Genetic engineering of mammalian cells is of interest as a means to boost bio-therapeutic protein yield. X-linked inhibitor of apoptosis (XIAP) overexpression has previously been shown to enhance CHO cell growth and prolong culture longevity while additionally boosting productivity. We confirmed this across a range of recombinant products (SEAP, EPO and IgG). However, stable over-expression of an engineering transgene competes for the cells translational machinery potentially compromising product titre. MicroRNAs are attractive genetic engineering candidates given their non-coding nature and ability to regulate multiple genes simultaneously, thereby relieving the translational burden associated with stable overexpression of a protein-encoding gene...
October 4, 2017: Biotechnology Journal
Prashant Sharma, Hyun Ah Cho, Jae-Won Lee, Woo Seung Ham, Bum Chul Park, Nam-Hyuk Cho, Young Keun Kim
Zinc oxide (ZnO) nanocomposites have been widely applied in biomedical fields due to their multifunctionality and biocompatibility. However, the physicochemical properties of ZnO nanocomposite involved in nano-bio interactions are poorly defined. To assess the potential applicability of ZnO nanowires for intracellular delivery of biomolecules, we examined the dynamics of cellular activity of cells growing on densely packed ZnO nanowire arrays with two different physical conformations, vertical (VNW) or fan-shaped (FNW) nanowires...
October 4, 2017: Nanoscale
Larissa S Mendes, Sybele Saska, Fernanda Coelho, Ticiana Sidorenko de Oliveira Capote, Raquel Mantuaneli Scarel-Caminaga, Reinaldo Marquetto, Raúl García Carrodeguas, Ana Maria Minarelli Gaspar, Miguel Angel Rodríguez
Calcium phosphate cements (CPC) have been widely investigated as bone graft substitutes due to its excellent self-setting ability, biocompatibility, osteoconductivity and moldability. In additional, mesoporous materials have also been studied as potential materials for application in medical devices due to their large surface area, which is capable of loading numerous biological molecules, besides being bioactive. In this study, bone β-TCP-MCPM-based injectable cement with mesoporous silica particles was synthesized and characterized in terms of their mechanical properties, microstructure, porosity, injectability, in vitro bioactivity and degradability; moreover, toxicity effects in CHO-K1 cells culture...
October 3, 2017: Biomedical Materials
Ho-Joon Lee, Myung Jin Son, Jiwon Ahn, Soo Jin Oh, Mihee Lee, Ansoon Kim, Yun-Ji Jeung, Han-Gyeul Kim, Misun Won, Jung Hwa Lim, Nam-Soon Kim, Cho-Rock Jung, Kyung-Sook Chung
Current in vitro liver models provide three-dimensional (3-D) microenvironments in combination with tissue engineering technology and can perform more accurate in vivo mimicry than two-dimensional models. However, a human cell-based, functionally mature liver model is still desired, which would provide an alternative to animal experiments and resolve low-prediction issues on species differences. Here, we prepared hybrid hydrogels of varying elasticity and compared them with a normal liver, to develop a more mature liver model that preserves liver properties in vitro...
September 28, 2017: Acta Biomaterialia
Tomoharu Hogiri, Hiroshi Tamashima, Akitoshi Nishizawa, Masahiro Okamoto
To optimize monoclonal antibody (mAb) production in Chinese hamster ovary cell cultures, culture pH should be temporally controlled with high resolution. In this study, we propose a new pH-dependent dynamic model represented by simultaneous differential equations including a minimum of six system component, depending on pH value. All kinetic parameters in the dynamic model were estimated using an evolutionary numerical optimization (real-coded genetic algorithm) method based on experimental time-course data obtained at different pH values ranging from 6...
September 27, 2017: Journal of Bioscience and Bioengineering
Yukako Senga, Hiroshi Imamura, Takamitsu Miyafusa, Hideki Watanabe, Shinya Honda
Therapeutic IgG becomes unstable under various stresses in the manufacturing process. The resulting non-native IgG molecules tend to associate with each other and form aggregates. Because such aggregates not only decrease the pharmacological effect but also become a potential risk factor for immunogenicity, rapid analysis of aggregation is required for quality control of therapeutic IgG. In this study, we developed a homogeneous assay using AlphaScreen and AF.2A1. AF.2A1 is a 25-residue artificial protein that binds specifically to non-native IgG generated under chemical and physical stresses...
September 29, 2017: Scientific Reports
Govindan Shanmugam, Amirthalingam Mohankumar, Duraisamy Kalaiselvi, Sundararaj Nivitha, Easwaran Murugesh, Piramanayagam Shanmughavel, Palanisamy Sundararaj
Caenorhabditis elegans is a sterol auxotroph requires minute amount of exogenous sterol for their growth and development. To culture the C. elegans, cholesterol was given as sterol molecule to maintain the optimum survival of worms. Diosgenin (DG), a plant derived steroidal saponin, structurally similar to cholesterol has been used as a precursor for the synthesis of steroidal hormones. In this study, worms were cultured with cholesterol (Cho(+)) and cholesterol-free (Cho(-)) medium with DG (5, 10 and 50μg/mL) at 20°C...
September 23, 2017: Biomedicine & Pharmacotherapy, Biomédecine & Pharmacothérapie
Lars Radke, Grit Sandig, Annika Lubitz, Ulrike Schließer, Hans Henning von Horsten, Veronique Blanchard, Karolin Keil, Volker Sandig, Christoph Giese, Michael Hummel, Stephan Hinderlich, Marcus Frohme
Subunit vaccines often require adjuvants to elicit sustained immune activity. Here, a method is described to evaluate the efficacy of single vaccine candidates in the preclinical stage based on cytokine and gene expression analysis. As a model, the recombinant human respiratory syncytial virus (RSV) fusion protein (RSV-F) was produced in CHO cells. For comparison, wild-type and glycoengineered, afucosylated RSV-F were established. Both glycoprotein vaccines were tested in a commercial Human Artificial Lymph Node in vitro model (HuALN(®))...
August 15, 2017: Bioengineering
Karen Schwab, Friedemann Hesse
Multi-wavelength fluorescence spectroscopy was evaluated in this work as tool for real-time monitoring of antibody aggregation in CHO fed-batch cultivations via partial least square (PLS) modeling. Therefore, we used the extrinsic fluorescence dyes 1-anilinonaphthalene-8-sulfonate (ANS), 4,4'-bis-1-anilinonaphthalene-8-sulfonate (Bis-ANS), or Thioflavin T (ThT) as medium additives. This is a new application area, since these dyes are commonly used for aggregate detection during formulation development. We determined the half maximum inhibitory concentrations of ANS (203 ± 11 µmol·L(-1)), Bis-ANS (5 ± 0...
July 24, 2017: Bioengineering
Xiao Pan, Ciska Dalm, René H Wijffels, Dirk E Martens
Normally, the growth profile of a CHO cell fed-batch process can be divided into two main phases based on changes in cell concentration, being an exponential growth phase and a stationary (non-growth) phase. In this study, an additional phase is observed during which the cell division comes to a halt but the cell growth continues in the form of an increase in cell size. The cell size increase (SI) phase occurs between the exponential proliferation phase (also called the number increase or NI phase) and the stationary phase...
September 26, 2017: Applied Microbiology and Biotechnology
Jeong Hwa Kim, Yeong-Jin Choi, Hee-Gyeong Yi, Joon Ho Wang, Dong-Woo Cho, Young Hun Jeong
Ligament, a fibrous connective tissue between bones, is a unique tissue in human anatomy because it has complex viscoelastic properties and is very tough. Moreover, it is an important tissue for regeneration because frequent injuries occur, but there are limited types of substitutes that can be used as a tissue replacement. In this study, we present a stem cell-laden fiber/hydrogel composite structure with a layered fibrous structure, which can enhance cell infiltration, topographical cue and mechanical properties...
September 25, 2017: Biomedical Materials
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