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"Culture Media"
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Reproducibility: Respect your cells
Vendors sell one serum-free medium for, say, Chinese hamster ovary cells, an epithelial cell line that is often used to produce therapeutic proteins, and others to expand particular types of blood cells. Concrete data, like good microscope images or expression data, can help researchers recognize when the cells used in their experiment have changed, says Anne Plant, a division chief at the US National Institute of Standards and Technology in Gaithersburg, Maryland, who hopes to find quantitative ways of making cellculture experiments comparable across laboratories.
Journal Article
Long-term ex vivo haematopoietic-stem-cell expansion allows nonconditioned transplantation
Multipotent self-renewing haematopoietic stem cells (HSCs) regenerate the adult blood system after transplantation
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, which is a curative therapy for numerous diseases including immunodeficiencies and leukaemias
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. Although substantial effort has been applied to identifying HSC maintenance factors through the characterization of the in vivo bone-marrow HSC microenvironment or niche
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–
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, stable ex vivo HSC expansion has previously been unattainable
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,
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. Here we describe the development of a defined, albumin-free culture system that supports the long-term ex vivo expansion of functional mouse HSCs. We used a systematic optimization approach, and found that high levels of thrombopoietin synergize with low levels of stem-cell factor and fibronectin to sustain HSC self-renewal. Serum albumin has long been recognized as a major source of biological contaminants in HSC cultures
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; we identify polyvinyl alcohol as a functionally superior replacement for serum albumin that is compatible with good manufacturing practice. These conditions afford between 236- and 899-fold expansions of functional HSCs over 1 month, although analysis of clonally derived cultures suggests that there is considerable heterogeneity in the self-renewal capacity of HSCs ex vivo. Using this system, HSC cultures that are derived from only 50 cells robustly engraft in recipient mice without the normal requirement for toxic pre-conditioning (for example, radiation), which may be relevant for HSC transplantation in humans. These findings therefore have important implications for both basic HSC research and clinical haematology.
An albumin-free culture system for the long-term ex vivo expansion of mouse haematopoietic stem cells produces 236- to 899-fold expansion, and generates cultures that robustly engraft in recipient mice without toxic pre-conditioning.
Journal Article
Bacterial nanocellulose production and application: a 10-year overview
Production of bacterial nanocellulose (BNC) is becoming increasingly popular owing to its environmentally friendly properties. Based on this benefit of BNC production, researchers have also begun to examine the capacity for cellulose production through microbial hosts. Indeed, several research groups have developed processes for BNC production, and many studies have been published to date, with the goal of developing methods for large-scale production. During BNC bioproduction, the culture medium represents approximately 30 % of the total cost. Therefore, one important and challenging aspect of the fermentation process is identification of a new cost-effective culture medium that can facilitate the production of high yields within short periods of time, thereby improving BNC production and permitting application of BNC in the biotechnological, medical, pharmaceutical, and food industries. In this review, we addressed different aspects of BNC production, including types of fermentation processes and culture media, with the aim of demonstrating the importance of these parameters.
Journal Article
Media Spectacle
During the mid-1990s, the O.J. Simpson murder trial dominated the media in the United States and were circulated throughout the world via global communications networks. The case became a spectacle of race, gender, class and violence, bringing in elements of domestic melodrama, crime drama and legal drama. According to this fascinating new book, the Simpson case was just one example of what the author calls 'media spectacle' - a form of media culture that puts contemporary dreams, nightmares, fantasies and values on display. Through the analysis of several such media spectacles - including Elvis, The X Files, Michael Jordan, and the Bill Clinton sex scandals - Doug Kellner draws out important insights into media, journalism, the public sphere and politics in an era of new technologies.
In this excellent follow up to his best selling Media Culture, Kellner's fascinating new volume delivers an informative read for students of sociology, culture and media.
eBook
Metabolic determinants of cancer cell sensitivity to glucose limitation and biguanides
New apparatus is used to maintain proliferating cancer cells in low-glucose conditions, demonstrating that mitochondrial oxidative phosphorylation (OXPHOS) is essential for optimal proliferation in these conditions; the most sensitive cell lines are defective in OXPHOS upregulation and may therefore be sensitive to current antidiabetic drugs that inhibit OXPHOS.
Biguanides active against starved tumour cells
Using a new continuous-flow culture apparatus called Nutrostat, designed to ensure constant and controlled extracellular nutrient levels, David Sabatini and colleagues screened cancer cell lines for genes important when cells experience low glucose levels. They found that the ability of cells to increase mitochondrial oxidative phosphorylation under conditions of low glucose was crucial. Cancer cells unable to do so due to impaired glucose utilization or mitochondrial DNA mutations were particularly sensitive to a class of compounds, biguanides, which are in use to treat diabetes. These findings may lead to new therapeutic applications of these drugs to treat tumours displaying such defects.
As the concentrations of highly consumed nutrients, particularly glucose, are generally lower in tumours than in normal tissues
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,
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, cancer cells must adapt their metabolism to the tumour microenvironment. A better understanding of these adaptations might reveal cancer cell liabilities that can be exploited for therapeutic benefit. Here we developed a continuous-flow culture apparatus (Nutrostat) for maintaining proliferating cells in low-nutrient media for long periods of time, and used it to undertake competitive proliferation assays on a pooled collection of barcoded cancer cell lines cultured in low-glucose conditions. Sensitivity to low glucose varies amongst cell lines, and an RNA interference (RNAi) screen pinpointed mitochondrial oxidative phosphorylation (OXPHOS) as the major pathway required for optimal proliferation in low glucose. We found that cell lines most sensitive to low glucose are defective in the OXPHOS upregulation that is normally caused by glucose limitation as a result of either mitochondrial DNA (mtDNA) mutations in complex I genes or impaired glucose utilization. These defects predict sensitivity to biguanides, antidiabetic drugs that inhibit OXPHOS
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,
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, when cancer cells are grown in low glucose or as tumour xenografts. Notably, the biguanide sensitivity of cancer cells with mtDNA mutations was reversed by ectopic expression of yeast NDI1, a ubiquinone oxidoreductase that allows bypass of complex I function
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. Thus, we conclude that mtDNA mutations and impaired glucose utilization are potential biomarkers for identifying tumours with increased sensitivity to OXPHOS inhibitors.
Journal Article