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https://www.readbyqxmd.com/read/28076798/neonatal-transplantation-confers-maturation-of-psc-derived-cardiomyocytes-conducive-to-modeling-cardiomyopathy
#1
Gun-Sik Cho, Dong I Lee, Emmanouil Tampakakis, Sean Murphy, Peter Andersen, Hideki Uosaki, Stephen Chelko, Khalid Chakir, Ingie Hong, Kinya Seo, Huei-Sheng Vincent Chen, Xiongwen Chen, Cristina Basso, Steven R Houser, Gordon F Tomaselli, Brian O'Rourke, Daniel P Judge, David A Kass, Chulan Kwon
Pluripotent stem cells (PSCs) offer unprecedented opportunities for disease modeling and personalized medicine. However, PSC-derived cells exhibit fetal-like characteristics and remain immature in a dish. This has emerged as a major obstacle for their application for late-onset diseases. We previously showed that there is a neonatal arrest of long-term cultured PSC-derived cardiomyocytes (PSC-CMs). Here, we demonstrate that PSC-CMs mature into adult CMs when transplanted into neonatal hearts. PSC-CMs became similar to adult CMs in morphology, structure, and function within a month of transplantation into rats...
January 10, 2017: Cell Reports
https://www.readbyqxmd.com/read/28068467/exodisc-for-rapid-size-selective-and-efficient-isolation-and-analysis-of-nanoscale-extracellular-vesicles-from-biological-samples
#2
Hyun-Kyung Woo, Vijaya Sunkara, Juhee Park, Tae-Hyeong Kim, Ja-Ryoung Han, Chi-Ju Kim, Hyun-Il Choi, Yoon-Keun Kim, Yoon-Kyoung Cho
Extracellular vesicles (EVs) are cell-derived, nanoscale vesicles that carry nucleic acids and proteins from their cells of origin and show great potential as biomarkers for many diseases, including cancer. Efficient isolation and detection methods are prerequisites for exploiting their use in clinical settings and understanding their physiological functions. Here, we presented a rapid, label-free, and highly sensitive method for EV isolation and quantification using a lab-on-a-disc integrated with two nanofilters (Exodisc)...
January 9, 2017: ACS Nano
https://www.readbyqxmd.com/read/28064070/maspin-inhibits-macrophage-phagocytosis-and-enhances-inflammatory-cytokine-production-via-activation-of-nf-%C3%AE%C2%BAb-signaling
#3
Yimeng Wang, Luguo Sun, Zhenbo Song, Danfeng Wang, Yongli Bao, Yuxin Li
Maspin (mammary serine protease inhibitor) is a non-inhibitory member of the serine protease inhibitor superfamily and a tumor suppressor in several cancers due to its ability to inhibit cell invasion, angiogenesis, and promote apoptosis. However, its immunomodulatory function remains largely unexplored. Thus, we explored the potential link between Maspin and macrophage function, first evaluating the regulatory effects of conditioned medium (CM) of a Maspin-overexpressing CHO cell strain on mouse peritoneal macrophage phagocytosis and cytokine secretion...
January 5, 2017: Molecular Immunology
https://www.readbyqxmd.com/read/28054753/t-helper-2-cytokine-induced-heat-shock-protein-70-secretion-and-its-potential-association-with-allergic-rhinitis
#4
Hyun Jin Min, Kyung Soo Kim, Joo-Heon Yoon, Chang-Hoon Kim, Hyung-Ju Cho
BACKGROUND: Various inflammatory mediators have been found to be involved in the pathogenesis of allergic disease (AR). The role of heat shock proteins in AR has not been studied. The aim of this study was to investigate the levels of heat shock protein 70 (Hsp70) in the nasal lavage fluids of AR patients and controls to elucidate the role of Hsp70 in the pathogenesis of AR. METHODS: Using an enzyme-linked immunosorbent assay, the levels of Hsp70, Hsp90, interleukin (IL)-4, IL-13, and IL-8 in nasal lavage fluid from patients were measured and statistically analyzed...
January 5, 2017: International Forum of Allergy & Rhinology
https://www.readbyqxmd.com/read/28054185/characterization-of-human-hybrid-cell-line-f2n78-through-a-comparison-of-culture-performances-and-protein-qualities
#5
Joon Serk Seo, Byung Sub Min, Yeon Jung Kim, Jong Moon Cho, Gi-Seong Kwon, Byeong-Pil Lim, Shin-Jae Chang, Dong-Il Kim
OBJECTIVES: To evaluate the characteristics of a novel human cell line, F2N78, including growth performance, physicochemical properties, and biological activity via direct comparison with CHO cells. RESULTS: The culture performance and physicochemical properties of antibodies produced from F2N78 and CHO cells were compared. For charge variants, antibodies produced from F2N78 cells contained a greater acidic charge variants than CHO cells. Regarding main glycoforms, degree of galactosylation was 52% in CT-A produced from F2N78 cells compared to CHO cells (37%)...
January 4, 2017: Biotechnology Letters
https://www.readbyqxmd.com/read/28045643/production-of-human-mutant-biologically-active-hepatocyte-growth-factor-in-chinese-hamster-ovary-cells
#6
Dongsheng Xu, Aini Wan, Lin Peng, Yun Chen, Yang He, Jianfeng Yang, Jian Jin
Hepatocyte growth factor (HGF) is a potent multi-functional cytokine that affects proliferation, migration and morphogenesis of various cells. HGF is secreted as an inactive single chain precursor protein and activated by the cleavage of serine proteases to form heterodimers. In our current study, the cleavage site of HGF was blocked by replaced Arg 494 of Glu (R494E) that resulted in the single chain HGF (R494E) unable to be cleaved by serine proteases. We established Chinese hamster ovary (CHO) cells overexpressing HGF (R494E), the expression of HGF (R494E) achieved 12 mg/L and was similar to a previously reported study...
January 3, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28042143/mesenchymal-stem-cells-ameliorate-b-cell-mediated-immune-responses-and-increase-il-10-expressing-regulatory-b-cells-in-an-ebi3-dependent-manner
#7
Kyung-Ah Cho, Jun-Kyu Lee, Yu-Hee Kim, Minhwa Park, So-Youn Woo, Kyung-Ha Ryu
Effector B cells are central contributors to the development of autoimmune disease by activating autoreactive T cells, producing pro-inflammatory cytokines and organizing ectopic lymphoid tissue. Conversely, IL-10-producing regulatory B (Breg) cells have pivotal roles in maintaining immunological tolerance and restraining excessive inflammation in autoinflammatory disease. Thus, regulating the equilibrium between antibody-producing effector B cells and Breg cells is critical for the treatment of autoimmune disease...
January 2, 2017: Cellular & Molecular Immunology
https://www.readbyqxmd.com/read/28034456/biotransformation-of-an-africanane-sesquiterpene-by-the-fungus-mucor-plumbeus
#8
Braulio M Fraga, Carmen E Díaz, Leonardo J Amador, Matías Reina, Matías López-Rodriguez, Azucena González-Coloma
Biotransformation of 8β-hydroxy-african-4(5)-en-3-one angelate by the fungus Mucor plumbeus afforded as main products 6α,8β-dihydroxy-african-4(5)-en-3-one 8β-angelate and 1α,8β-dihydroxy-african-4(5)-en-3-one 8β-angelate, which had been obtained, together with the substrate, from transformed root cultures of Bethencourtia hermosae. This fact shows that the enzyme system involved in these hydroxylations in both organisms, the fungus and the plant, acts with the same regio- and stereospecificity. In addition another twelve derivatives were isolated in the incubation of the substrate, which were identified as the (2'R,3'R)- and (2'S,3'S)-epoxy derivatives of the substrate and of the 6α- and 1α-hydroxy alcohols, the 8β-(2'R,3'R)- and 8β-(2'S,3'S)-epoxyangelate of 8β,15-dihydroxy-african-4(5)-en-3-one, the hydrolysis product of the substrate, and three isomers of 8β-hydroxy-african-4(5)-en-3-one 2ξ,3ξ-dihydroxy-2-methylbutanoate...
December 26, 2016: Phytochemistry
https://www.readbyqxmd.com/read/28029182/nedd4-deficiency-in-vascular-smooth-muscle-promotes-vascular-calcification-by-stabilizing-psmad1
#9
Ji-Hyun Lee, Seon-Ae Jeon, Byung-Gyu Kim, Michiko Takeda, Jae-Jin Cho, Dong-Ik Kim, Hiroshi Kawabe, Je-Yoel Cho
The nonosseous calcification process such as atherosclerosis is one of the major complications in several types of metabolic diseases. In previous study, we uncovered that aberrant activity of TGF-β signaling pathway could contribute the vascular smooth muscle cells (VSMCs) calcification process. Also, we identified NEDD4 E3 ligase as a key suppressor of BMP/Smad pathway via a polyubiquitination dependent selective degradation of C-terminal phosphorylated Smad1 (pSmad1) activated by TGF-β⋅ Here, we further validated and confirmed the role of Nedd4 in in vivo vascular calcification progression...
December 28, 2016: Journal of Bone and Mineral Research: the Official Journal of the American Society for Bone and Mineral Research
https://www.readbyqxmd.com/read/28025159/anti-apoptotic-genes-bcl-2-and-bcl-xl-overexpression-can-block-iridovirus-serine-threonine-kinase-induced-bax-mitochondria-mediated-cell-death-in-gf-1%C3%A2-cells
#10
Latif Reshi, Hua-Ven Wang, Cho-Fat Hui, Yu-Chin Su, Jiann-Ruey Hong
Although serine/threonine (ST) kinase is known to induce host cell death in GF-1 cells, it remains unclear how ST kinase induces mitochondrial function loss. In the present study, we addressed the issue of mitochondrial function loss by determining whether the Bcl-2 family members Bcl-2 and Bcl-xL can prevent ST kinase-induced cell death activity via interacting with the pro-apoptotic gene Bax. Grouper fin cells (GF-1) carrying EGFP-Bal-xL and EGFP-Bcl-2 fused genes were selected, established in cell culture, and used to examine the involvement of Bcl-2 and Bcl-xL overexpression in protection of GF-1 cells from the effects of the giant sea perch iridovirus (GSIV) ST kinase gene...
December 23, 2016: Fish & Shellfish Immunology
https://www.readbyqxmd.com/read/28012362/quantification-and-simultaneous-evaluation-of-the-bioactivity-of-antibody-produced-in-cho-cell-culture-the-use-of-the-ectodomain-of-fc%C3%AE-ri-and-surface-plasmon-resonance-based-biosensor
#11
July Dorion-Thibaudeau, Yves Durocher, Gregory De Crescenzo
A surface plasmon resonance (SPR)-based assay has been developed in order to quantify the monoclonal antibody (Mab) Trastuzumab within the supernatant of a mammalian cell culture using the ectodomain of FcγRI (CD64) and confirm Mab bioactivity, i.e. binding to its antigen Her2, in a single biosensing experiment. Under partial mass transport limitation, we were able to quantify Mab present in unpurified samples taken throughout the cell culture. While Mab capture on the biosensor surface confirmed the ability of its Fc region to bind to FcγRI, the binding activity of its Fab region was also tested by injecting increasing concentrations of the Mab antigen (Her2)...
December 21, 2016: Molecular Immunology
https://www.readbyqxmd.com/read/28003219/direct-reprogramming-of-human-dermal-fibroblasts-into-endothelial-cells-using-er71-etv2
#12
Sangho Lee, Changwon Park, Ji W Han, Ju Young Kim, Kyuwon Cho, Eun Jae Kim, Sangsung Kim, Shin-Jeong Lee, Se-Yeong Oh, Yoshiaki Tanaka, In-Hyun Park, Hyo Jae An, Claire M Shin, Shraya Sharma, Young-Sup Yoon
RATIONALE: Direct conversion or reprogramming of human postnatal cells into endothelial cells (ECs), bypassing stem or progenitor cell status, is crucial for regenerative medicine, cell therapy, and pathophysiological investigation but has remained largely unexplored. OBJECTIVE: We sought to directly reprogram human postnatal dermal fibroblasts (HDFs) to ECs with vasculogenic and endothelial transcription factors (TFs) and determine their vascularizing and therapeutic potential...
December 21, 2016: Circulation Research
https://www.readbyqxmd.com/read/27997765/intracellular-cho-cell-metabolite-profiling-reveals-steady-state-dependent-metabolic-fingerprints-in-perfusion-culture
#13
Daniel J Karst, Robert Steinhoff, Marie R G Kopp, Elisa Serra, Miroslav Soos, Renato Zenobi, Massimo Morbidelli
Perfusion cell culture processes allow the steady-state culture of mammalian cells at high viable cell density, which is beneficial for overall product yields and homogeneity of product quality in the manufacturing of therapeutic proteins. In this study, the extent of metabolic steady state and the change of the metabolite profile between different steady states of an industrial Chinese hamster ovary (CHO) cell line producing a monoclonal antibody (mAb) was investigated in stirred tank perfusion bioreactors...
December 20, 2016: Biotechnology Progress
https://www.readbyqxmd.com/read/27986089/the-anti-inflammatory-role-of-extranuclear-apurinic-apyrimidinic-endonuclease-1-redox-effector-factor-1-in-reactive-astrocytes
#14
Hyunjung Baek, Chae Seong Lim, Hee Sun Byun, Hyun Sil Cho, Yu Ran Lee, Yong Sup Shin, Hyun-Woo Kim, Byeong Hwa Jeon, Dong Woon Kim, Jinpyo Hong, Gang Min Hur, Jin Bong Park
Apurinic/apyrimidinic endonuclease 1 (APE1), a ubiquitous multipurpose protein, is also known as redox effector factor-1 (Ref-1). It is involved in DNA repair and redox signaling and, in turn, oxidative stress-induced neurodegeneration. Although previous studies have demonstrated that APE1/Ref-1 functions as a negative regulator of inflammatory response via several mechanisms in neuronal cells, little is known about the roles of APE1/Ref-1 in glial cells. In this study, we found that cytoplasmic APE1/Ref-1 expression was upregulated in reactive astrocytes of the kainic acid- or lipopolysaccharide (LPS)-injected hippocampus...
December 16, 2016: Molecular Brain
https://www.readbyqxmd.com/read/27983869/pharmacological-rescue-of-conditionally-reprogrammed-cystic-fibrosis-bronchial-epithelial-cells
#15
Martina Gentzsch, Susan E Boyles, Chaitra Cheluvaraju, Imron G Chaudhry, Nancy L Quinney, Crescentia Cho, Hong Dang, Xuefeng Liu, Richard Schlegel, Scott H Randell
Well-differentiated primary human bronchial epithelial (HBE) cell cultures are vital for cystic fibrosis (CF) research, particularly for development of CF transmembrane conductance regulator (CFTR) modifier drugs. Culture of epithelial cells with irradiated 3T3 fibroblast feeder cells plus the RhoA kinase (ROCK) inhibitor Y-27632 (Y), termed conditionally reprogrammed cell (CRC) technology, enhances cell growth and lifespan while preserving cell of origin functionality. We initially determined the electrophysiological and morphological characteristics of conventional versus CRC expanded non-CF HBE cells...
December 16, 2016: American Journal of Respiratory Cell and Molecular Biology
https://www.readbyqxmd.com/read/27977914/analysis-of-tubespins-as-a-suitable-scale-down-model-of-bioreactors-for-high-cell-density-cho-cell-culture
#16
Natalia Gomez, Malhar Ambhaikar, Li Zhang, Chung-Jr Huang, Hedieh Barkhordarian, Jonathan Lull, Chris Gutierrez
High cell density (HCD) culture increases recombinant protein productivity via higher biomass. Compared to traditional fed-batch cultures, HCD is achieved by increased nutrient availability and removal of undesired metabolic components via regular medium replenishment. HCD process development is usually performed in instrumented lab-scale bioreactors (BR) that require time and labor for set-up and operation. To potentially minimize resources and cost during HCD experiments, we evaluated a two-week 50mL Tubespin (TS) simulated HCD process where daily medium exchanges mimic the medium replacement rate in BR...
December 15, 2016: Biotechnology Progress
https://www.readbyqxmd.com/read/27977910/bioreactor-productivity-and-media-cost-comparison-for-different-intensified-cell-culture-processes
#17
Sen Xu, John Gavin, Rubin Jiang, Hao Chen
Process intensification in biomanufacturing has attracted a great deal of interest in recent years. Manufacturing platform improvements leading to higher cell density and bioreactor productivity have been pursued. Here we evaluated a variety of intensified mammalian cell culture processes for producing monoclonal antibodies. Cell culture operational modes including fed-batch (normal seeding density or high seeding density with N-1 perfusion), perfusion, and concentrated fed-batch (CFB) were assessed using the same media set with the same Chinese Hamster Ovary (CHO) cell line...
December 15, 2016: Biotechnology Progress
https://www.readbyqxmd.com/read/27943242/knockout-of-a-difficult-to-remove-cho-host-cell-protein-lipoprotein-lipase-for-improved-polysorbate-stability-in-monoclonal-antibody-formulations
#18
Josephine Chiu, Kristin N Valente, Nicholas E Levy, Lie Min, Abraham M Lenhoff, Kelvin H Lee
While the majority of host cell protein (HCP) impurities are effectively removed in typical downstream purification processes, a small population of HCPs are particularly challenging. Previous studies have identified HCPs that are challenging for a variety of reasons. Lipoprotein lipase (LPL)-a Chinese hamster ovary (CHO) HCP that functions to hydrolyze esters in triglycerides-was one of ten HCPs identified in previous studies as being susceptible to retention in downstream processing. LPL may degrade polysorbate 80 (PS-80) and polysorbate 20 (PS-20) in final product formulations due to the structural similarity between polysorbates and triglycerides...
December 12, 2016: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/27941997/primary-cilia-negatively-regulate-melanogenesis-in-melanocytes-and-pigmentation-in-a-human-skin-model
#19
Hyunjung Choi, Ji Hyun Shin, Eun Sung Kim, So Jung Park, Il-Hong Bae, Yoon Kyung Jo, In Young Jeong, Hyoung-June Kim, Youngjin Lee, Hea Chul Park, Hong Bae Jeon, Ki Woo Kim, Tae Ryong Lee, Dong-Hyung Cho
The primary cilium is an organelle protruding from the cell body that senses external stimuli including chemical, mechanical, light, osmotic, fluid flow, and gravitational signals. Skin is always exposed to the external environment and responds to external stimuli. Therefore, it is possible that primary cilia have an important role in skin. Ciliogenesis was reported to be involved in developmental processes in skin, such as keratinocyte differentiation and hair formation. However, the relation between skin pigmentation and primary cilia is largely unknown...
2016: PloS One
https://www.readbyqxmd.com/read/27929401/electrical-impedance-monitoring-of-c2c12-myoblast-differentiation-on-an-indium-tin-oxide-electrode
#20
Ilhwan Park, Yeonhee Hong, Young-Hoo Jun, Ga-Yeon Lee, Hee-Sook Jun, Jae-Chul Pyun, Jeong-Woo Choi, Sungbo Cho
Electrical cell-substrate impedance sensing is increasingly being used for label-free and real-time monitoring of changes in cell morphology and number during cell growth, drug screening, and differentiation. In this study, we evaluated the feasibility of using ECIS to monitor C2C12 myoblast differentiation using a fabricated indium tin oxide (ITO) electrode-based chip. C2C12 myoblast differentiation on the ITO electrode was validated based on decreases in the mRNA level of MyoD and increases in the mRNA levels of myogenin and myosin heavy chain (MHC)...
December 5, 2016: Sensors
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