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Articles by Maria Rende in JoVE
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Ex vivo Mimicry den normala respektive onormala mänskliga Hematopoiesis
Teresa Mortera-Blanco1, Maria Rende1, Hugo Macedo1, Serene Farah1, Alexander Bismarck1, Athanasios Mantalaris1, Nicki Panoskaltsis2
1Department of Chemical Engineering and Chemical Technology, South Kensington campus, Imperial College London, 2Department of Hematology, Northwick Park & St. Mark's campus, Imperial College London
En 3D-odlingssystem för hematopoies beskrivs med humant navelsträngsblod och leukemiska benmärgsceller. Metoden baseras på användningen av en porös syntetisk polyuretan byggnadsställning belagd med extracellulära matrisproteiner. Detta ställningen är anpassningsbar för att rymma en mängd olika celler.
Other articles by Maria Rende on PubMed
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Long-term Maintenance of Human Hepatocytes in Oxygen-permeable Membrane Bioreactor
Biomaterials.
Sep, 2006 |
Pubmed ID: 16753210 An oxygen-permeable membrane bioreactor utilizing human hepatocytes has been tested in this study. In the bioreactor, human hepatocytes were cultured between flat-sheet gas-permeable polymeric membranes, which ensure the diffusion of O(2) and CO(2) providing a support for cell anchorage and growth and permit the online observation of the cells with an inverse microscope. This bioreactor allows a direct oxygenation of cells adhered on membranes and of the medium overlaying cells simulating in vivo sinusoidal organization. Human hepatocytes were cultured in the presence of some therapeutic molecules to assess the temporal liver-specific functions of the cells. Interleukin 6 (IL-6), which is a multifactorial proinflammatory cytokine involved in a variety of host defences and pathological processes, and diclofenac, an arylacetic non-steroidal anti-inflammatory drug, were used as therapeutic molecules. The aim of this study was to evaluate the in vitro performance of the small oxygen-permeable membrane bioreactor in the long-term maintenance and differentiation of human hepatocytes under in-vivo-like conditions. The fluid dynamics of the bioreactor were characterized before using it for human cell culture. The functional response to a step challenge in the medium of IL-6 (120 pg/ml), diclofenac (80 microm) and IL-6 and diclofenac together was investigated. The ability of hepatocytes to perform liver-specific functions in terms of urea and albumin synthesis, as well as secretion of total proteins, was maintained for 32 days. Also, the diclofenac biotransformation functions were sustained as the formation of the metabolites 4'-OH-diclofenac and 5-OH-diclofenac lactam demonstrated. This study attested the feasibility of the membrane bioreactor as an in vitro simple model system that allows human hepatocytes to be maintained in a differentiated state similar to that in vivo.
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Human Lymphocyte PEEK-WC Hollow Fiber Membrane Bioreactor
Journal of Biotechnology.
Oct, 2007 |
Pubmed ID: 17905461 In this study we developed a PEEK-WC hollow fiber (HF) membrane bioreactor for the maintenance of human peripheral lymphocytes as model system for the in vitro investigation of disease pathogenesis, chemical effects and individual drug sensitivity. Peripheral lymphocytes isolated from donor's human buffy coat were cultured in the shell compartment of the PEEK-WC-HF bioreactor and stimulated with PHA 5microg/mL for the first 48h of culture to enhance cytokine production and cell proliferation. Thereafter, cells were cultured in the presence of Hypericum perforatum (St. John's wort) in order to induce cytochrome P450s enzymes, CYP2E, involved in the biotransformation of endogenous molecules and exogenous compounds. The metabolic activity of cells with respect to glucose consumption and oxygen uptake was maintained for all the culture time without the addition of mitogen. Two cytokines IL-2 and IL-10, which are specific pattern of lymphocytes T helper 1 and T helper 2, respectively, were produced in the bioreactor up to 14 days of culture. Lymphocytes were also able to biotransform acetaminophen through the formation of the main metabolite paracetamidofenil-beta-glucuronide, which is the product of glucuronidation reaction, as a result of the Hypericum perforatum administration that induced the catalytic activity of the CYP2E1. These results demonstrated the usefulness of the bioreactor as the support system that reproduces physiological parameters such as a constant perfusion of medium, nutrients and oxygen maintaining the in vitro integrity of lymphocyte viability and functions.
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Distinct Alpha Subunits of the GABAA Receptor Are Responsible for Early Hippocampal Silent Neuron-related Activities
Hippocampus.
Nov, 2009 |
Pubmed ID: 19338020 The modulatory actions of GABA(A) receptor subunits are crucial for morphological and transcriptional neuronal activities. In this study, growth of hamster hippocampal neurons on biohybrid membrane substrates allowed us to show for the first time that the two major GABA(A) alpha receptor subunits (alpha(2,5)) are capable of early neuronal shaping plus expression differences of some of the main neuronal cytoskeletal factors (GAP-43, the neurotrophin--BDNF) and of Gluergic subtypes. In a first case the inverse alpha(5) agonist (RY-080) seemed to account for the reduction of dendritic length at DIV7, very likely via lower BDNF levels. Conversely, the effects of the preferentially specific agonist for hippocampal alpha(2) subunit (flunitrazepam) were, instead, directed at the formation of growth cones at DIV3 in the presence of greatly (P < 0.01) diminished GAP-43 levels as displayed by strongly reduced axonal sprouting. It is interesting to note that concomitantly to these morphological variations, the transcription of some Gluergic receptor subtypes resulted to be altered. In particular, flunitrazepam was responsible for a distinctly rising expression of axonal NR1 mRNA levels from DIV3 (P < 0.01) until DIV7 (P < 0.001), whereas RY-080 evoked a very great (P < 0.001) downregulation of dendritic GluR2 at only DIV7. Together, our results demonstrate that GABA(A) alpha(2,5) receptor-containing subunits by regulating the precise synchronization of cytoskeletal factors are considered key modulating neuronal elements of hippocampal morphological growth features. Moreover, the notable NR1 and GluR2 transcription differences promoted by these GABA(A) alpha subunits tend to favorably corroborate the early role of alpha(2) + alpha(5) on hippocampal neuronal networks in hibernating rodents through the recruitment and activation of silent neurons, and this may provide useful insights regarding molecular neurodegenerative events.
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