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CHO cell culture

Fatemeh Torkashvand, Fereidoun Mahboudi, Manouchehr Vossoughi, Elnaz Fatemi, Seyed Masoud Moosavi Basri, Amir Heydari, Behrouz Vaziri
Background: Chinese hamster ovary (CHO) cell line is considered as the most common cell line in the biopharmaceutical industry because of its capability in performing efficient post-translational modifications and producing the recombinant proteins, which are similar to natural human proteins. The optimization of the upstream process via different feed strategies has a great impact on the target molecule expression and yield. Methods: To determine and understand the molecular events beneath the feed effects on the CHO cell, a label-free quantitative proteomic analysis was applied...
April 21, 2018: Iranian Biomedical Journal
Soon Ok Cho, Min-Hyun Kim, Hyeyoung Kim
Background: Oxidative stress occurs in white adipose tissue and dysregulates the expression of adipokines secreted from adipocytes. Since adipokines influence inflammation, supplementation with antioxidants might be beneficial for preventing oxidative stress-mediated inflammation in adipocytes and inflammation-associated complications. β-Carotene is the most prominent antioxidant carotenoid and scavenges reactive oxygen species in various tissues. The purpose of this study was to determine whether β-carotene regulates the expression of adipokines, such as adiponectin, monocyte chemoattractant protein-1 (MCP-1), and regulated on activation, normal T cell expressed and secreted (RANTES) in 3T3-L1 adipocytes treated with glucose/glucose oxidase (G/GO)...
March 2018: Journal of Cancer Prevention
Mi-Young Son, Cho-Rok Jung, Dae-Soo Kim, Hyun-Soo Cho
The technology of tissue differentiation from human pluripotent stem cells has attracted attention as a useful resource for regenerative medicine, disease modeling and drug development. Recent studies have suggested various key factors and specific culture methods to improve the successful tissue differentiation and efficient generation of human induced pluripotent stem cells. Among these methods, epigenetic regulation and epigenetic signatures are regarded as an important hurdle to overcome during reprogramming and differentiation...
April 6, 2018: Molecular Biology Reports
Andi Ariyandy, Chiemi Sakai, Mari Ishida, Ryusei Mizuta, Kiyoshi Miyagawa, Satoshi Tashiro, Aiko Kinomura, Koji Hiraaki, Keitaro Ueda, Masao Yoshizumi, Takafumi Ishida
Deficiency of X-ray repair cross-complementing protein 3 (XRCC3), a DNA-damage repair molecule, and the 241Met variant of XRCC3 have been reported to increase endoreduplication, which induces polyploidy. The aims of this study were to determine the impact of the XRCC3 polymorphism on the incidence of hypertension-induced left ventricular hypertrophy (LVH) and to investigate the mechanisms underlying any potential relationship. Patients undergoing chronic hemodialysis (n = 77) were genotyped to assess for the XRCC3 Thr241Met polymorphism...
April 6, 2018: Hypertension Research: Official Journal of the Japanese Society of Hypertension
Seul Ki Seo, Nari Kim, Ju Hee Lee, Sang Min Kim, Sang Yeub Lee, Jang Whan Bae, Kyung Kuk Hwang, Dong Woon Kim, Walter J Koch, Myeong Chan Cho
BACKGROUND AND OBJECTIVES: β-arrestin2 (β-arr2) basically regulates multiple signaling pathways in mammalian cells by desensitization and internalization of G-protein coupled receptors (GPCRs). We investigated impacts of β-arr2 on survival, mobility, and tube formation of cardiac progenitor cells (CPCs) obtained from wild-type (WT) mouse (CPC-WT), and β-arr2 knock-out (KO) mouse (CPC-KO) cultured in presence or absence of serum and oxygen as non-canonical roles in GPCR system. METHODS: CPCs were cultured in Dulbecco's Modified Eagle Medium/Nutrient Mixture F-12 -based media containing fetal bovine serum and growth factors...
April 2018: Korean Circulation Journal
William S Kish, Matthew K Roach, Hiroyuki Sachi, Amith D Naik, Stefano Menegatti, Ruben G Carbonell
Prior work described the identification and characterization of erythropoietin-binding cyclic peptides SLFFLH, VVFFVH, FSLLHH and FSLLSH (all of the form cyclo[(Nα -Ac)Dap(A)-X1 -X6 -AE], wherein X1 -X6 is the listed sequences). In this work, the peptide ligands were synthesized on Toyopearl chromatographic resins and utilized for purifying recombinant human erythropoietin (rHuEPO) from complex sources. Elution buffer pH and composition were optimized to maximize the recovery of standard rHuEPO from the peptide resins...
March 27, 2018: Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
Sangkyun Cho, Amal Abbas, Jerome Irianto, Irena Ivanovska, Yuntao Xia, Manu Tewari, Dennis E Discher
Interphase phosphorylation of lamin-A,C depends dynamically on a cell's microenvironment, including the stiffness of extracellular matrix. However, phosphorylation dynamics is poorly understood for diseased forms such as progerin, a permanently farnesylated mutant of LMNA that accelerates aging of stiff and mechanically stressed tissues. Here, fine-excision alignment mass spectrometry (FEA-MS) is developed to quantify progerin and its phosphorylation levels in patient iPS cells differentiated to mesenchymal stem cells (MSCs)...
April 5, 2018: Nucleus
Mahmood Rahimi-Zarchi, Seyed Abbas Shojaosadati, Mohammad Mehdi Amiri, Mahmood Jeddi-Tehrani, Fazel Shokri
The effects of all-trans retinoic acid (RA) and sodium butyrate (NaBu) on growth, viability and antibody production of two types of transfected Chinese hamster ovary cell lines (CHO-K1 and CHO-S) were investigated using a batch mode cell culture. By adding 0.5 mM NaBu in the CHO-K1 cell culture, the cell specific productivity (Qp ) and antibody concentration increased by five- and threefold, respectively. The optimal concentration of RA was 100 nM which resulted in twofold increase in antibody production...
April 4, 2018: Bioprocess and Biosystems Engineering
Byoung Soo Kim, Yang Woo Kwon, Jeong-Sik Kong, Gyu Tae Park, Ge Gao, Wonil Han, Moon-Bum Kim, Hyungseok Lee, Jae Ho Kim, Dong-Woo Cho
3D cell-printing technique has been under spotlight as an appealing biofabrication platform due to its ability to precisely pattern living cells in pre-defined spatial locations. In skin tissue engineering, a major remaining challenge is to seek for a suitable source of bioink capable of supporting and stimulating printed cells for tissue development. However, current bioinks for skin printing rely on homogeneous biomaterials, which has several shortcomings such as insufficient mechanical properties and recapitulation of microenvironment...
March 23, 2018: Biomaterials
Simone Albrecht, Christian Kaisermayer, David Reinhart, Monica Ambrose, Renate Kunert, Anna Lindeberg, Jonathan Bones
The monitoring of protein biomarkers for the early prediction of cell stress and death is a valuable tool for process characterization and efficient biomanufacturing control. A representative set of six proteins, namely GPDH, PRDX1, LGALS1, CFL1, TAGLN2 and MDH, which were identified in a previous CHO-K1 cell death model using discovery LC-MSE was translated into a targeted liquid chromatography multiple reaction monitoring mass spectrometry (LC-MRM-MS) platform and verified. The universality of the markers was confirmed in a cell growth model for which three Chinese hamster ovary host cell lines (CHO-K1, CHO-S, CHO-DG44) were grown in batch culture in two different types of basal media...
April 2, 2018: Analytical and Bioanalytical Chemistry
Thaddaeus A Webster, Brian C Hadley, William Hilliard, Colin Jaques, Carrie Mason
The monitoring and control of bioprocesses is of the utmost importance in order to provide a consistent, safe, and high quality product for consumers. Current monitoring and control schemes rely on infrequent and time consuming offline sampling methods, which inherently leads to some variability in the process which may impact the product quality profile. As part of Lonza's dedication to process analytical technology (PAT) initiatives this study evaluated the ability to generate generic calibration models, which are independent of the cell line, using Raman probes to monitor changes in glucose, lactate, glutamate, ammonium, viable cell concentration (VCC), total cell concentration (TCC) and product concentration...
March 31, 2018: Biotechnology Progress
Fiona Hartley, Tracy Walker, Vicky Chung, Karl Morten
The metabolism of Chinese Hamster Ovary (CHO) cells in a production environment has been extensively investigated. However, a key metabolic transition, the switch from lactate production to lactate consumption, remains enigmatic. Though commonly observed in CHO cultures, the mechanism(s) by which this metabolic shift is triggered is unknown. Despite this, efforts to control the switch have emerged due to the association of lactate consumption with improved cell growth and productivity. This review aims to consolidate current theories surrounding the lactate switch...
March 31, 2018: Biotechnology and Bioengineering
Yuko Matsuda, Otomi Cho, Takashi Sugita, Daiki Ogishima, Satoru Takeda
PURPOSE: Vulvovaginal candidiasis (VVC) is a common superficial infection of the vaginal mucous membranes caused by the fungus Candida albicans. The aim of this study was to assess the mechanisms underlying the inhibitory effects of the culture supernatants of Lactobacillus gasseri and L. crispatus, the predominant microbiota in Asian healthy women, on C. albicans biofilm formation. The inhibition of C. albicans adhesion to HeLa cells by Lactobacillus culture supernatant was also investigated...
March 30, 2018: Mycopathologia
Nayoun Kim, Young-Sun Nam, Keon-Il Im, Jung-Yeon Lim, Young-Woo Jeon, Yunejin Song, Jong Wook Lee, Seok-Goo Cho
Background: Cytomegalovirus(CMV)-related diseases are a serious cause of morbidity and mortality following hematopoietic stem cell transplantation (HSCT). CMV-specific cytotoxic T lymphocytes (CMV-CTLs) have been reported as an alternative to antiviral drugs that provide long-term CMV-specific immunity without major side effects. However, their application has been limited by the prolonged manufacturing process required. Methods: In this study, we applied the IFN-γ cytokine capture system (CCS) using the fully automated CliniMACS Prodigy device for rapid production of CMV-CTLs, which may be applicable in clinically urgent CMV-related diseases...
January 2018: Transfusion Medicine and Hemotherapy
Shengwei Ji, Yuzi Luo, Tengteng Zhang, Lina Shao, Xing-Yu Meng, Yimin Wang, Yao Gao, Yongfeng Li, Su Li, Yuan Sun, Xin Jin, Hua-Ji Qiu
Classical swine fever (CSF), which is caused by classical swine fever virus (CSFV), is a highly contagious disease of pigs. CSFV is genetically and serologically related to bovine viral diarrhea virus (BVDV), a ruminant pestivirus. However, currently available ELISAs based on the full-length E2 protein of CSFV cannot discriminate anti-CSFV from anti-BVDV antibodies. In this study, a truncated CSFV E2 protein (amino acids 690 to 879) covering antigenic domains B/C/D/A (E2B/C/D/A ) was designed based on homologous modeling according to the crystal structure of the BVDV E2 protein...
March 23, 2018: Archives of Virology
Jae-Uk Jeong, Tung Nguyen Thanh Uong, Woong-Ki Chung, Taek-Keun Nam, Sung-Ja Ahn, Ju-Young Song, Sang-Ki Kim, Dong-Jun Shin, Eugene Cho, Kyoung Won Kim, Duck Cho, Mee Sun Yoon
BACKGROUND AIMS: Irradiation enhances the adhesion between natural killer (NK) cells and target cells by up-regulating intercellular adhesion molecule-1 (ICAM-1) on target cells. Therefore, we investigated the effect of irradiation-induced ICAM-1 expression on human cancer cells on NK cell-mediated cytotoxicity. METHODS: Expression levels of ICAM-1 on the target cell surface before and after irradiation of six human cancer cell lines (HL60, SKBR-3, T47D, HCT-116, U937 and U251) were analyzed by flow cytometry...
March 20, 2018: Cytotherapy
Mauro Torres, Roberto Zúñiga, Matias Gutierrez, Mauricio Vergara, Norberto Collazo, Juan Reyes, Julio Berrios, Juan Carlos Aguillon, Maria Carmen Molina, Claudia Altamirano
Chinese hamster ovary (CHO) cells are the most frequently used host for commercial production of therapeutic proteins. However, their low protein productivity in culture is the main hurdle to overcome. Mild hypothermia has been established as an effective strategy to enhance protein specific productivity, although the causes of such improvement still remain unclear. The self-regulation of global transcriptional regulatory factors, such as Myc and XBP1s, seems to be involved in increased the recombinant protein production at low temperature...
2018: PloS One
Hyun-Ook Jeon, In-Seok Choi, Ji-Young Yoon, Eun-Jung Kim, Ji-Uk Yoon, Ah-Reum Cho, Hyung-Joon Kim, Cheul-Hong Kim
Background: The structure and function of bone tissue is maintained through a constant remodeling process, which is maintained by the balance between osteoblasts and osteoclasts. The failure of bone remodeling can lead to pathological conditions of bone structure and function. Remifentanil is currently used as a narcotic analgesic agent in general anesthesia and sedation. However, the effect of remifentanil on osteoclasts has not been studied. Therefore, we investigated the effect of remifentanil on pre-osteoclast (pre-OCs) differentiation and the mechanism of osteoclast differentiation in the absence of specific stimulus...
February 2018: Journal of Dental Anesthesia and Pain Medicine
Hyeran Cho, Aeri Lee, Kyobum Kim
Background: Fetal bovine serum (FBS) is the most essential supplement in culture media for cellular proliferation, metabolism, and differentiation. However, due to a limited supply and subsequently rising prices, a series of studies have investigated a biological feasibility of replaceable serums to substitute FBS. Along with the increasing interests to manufacture stem cell-based cellular products, optimizing the composition of culture media including serums and exogenous growth factors (GFs) is of importance...
2018: Biomaterials Research
Seung Hyun Lee, Eun-Hye Seo, Hyun Jun Park, Chung-Sik Oh, Cho Long Kim, Sewon Park, Seong-Hyop Kim
This study assessed the effects of crystalloid versus synthetic colloid in vitro on immune cells co-cultured with mouse splenocytes. Mouse splenocytes were co-cultured with three different types of fluid: Plasma solution-A® (CJ HealthCare, Seoul, Korea; the crystalloid group); Tetraspan 6%® (B. Braun Medical, Melsungen, Germany; the Colloid-T group); and Volulyte 6%® (Fresenius Kabi, Bad Homburg vor dér-Höhe, Germany; Colloid-V group). To evaluate the acquired immune response, cluster of differentiation (CD) 4+ T cells and CD8+ T cells were measured...
March 19, 2018: Scientific Reports
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