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          kirkstall代理QV500, QV600, QV900,Quasi-Vivo &amp;#174; Systems,3D灌流培養(yǎng)系統(tǒng)

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           kirkstall代理QV500, QV600, QV900,Quasi-Vivo ® Systems,3D灌流培養(yǎng)系統(tǒng)

           

          Quasi-Vivo體外長(zhǎng)期流灌注細(xì)胞培養(yǎng)系統(tǒng)

          ——Long term cell-culture breakthrough

          Oview

          Kirkstall has recently launched the Quasi-Vivo® 500 Starter Kit for use by the research community. The dynamic nutrient flow means that unlike conventional static cell culture, cell to cell signalling is permitted between different cell and tissue types, whilst the bio-module design minimises the shear stress in the cell culture region, improving cell vitality and allowing longer duration experiments.

          800px-Minusheet_Figure_3.jpg

           

          THE SYSTEM FEATURES INCLUDE:

          Ø  Dynamic flow of nutrient media through multiple culture

          Ø  chambers.

          Ø  Media and flow rates can be selected to allow successful long

          Ø  term culture of different cell types in the same experiment.

          Ø  Can be used with many standard CO2 incubators by mounting

          Ø  Quasi-Vivo® components on a convenient tray.

          Ø  Modular construction enables chambers to be connected in

          Ø  series or parallel.

          Ø  The key feature of Quasi-Vivo® systems is the ability to culture cells under flow.

           

          Each Kit Comprises:

          Ø   3 bioreactor chambers 1.5 ml capacity

          Ø   Connecting tubes 1.5 mm internal diameter

          Ø   Tube connectors

          Ø  100 glass coverslips 12 mm diameter

          Ø   1 Reservoir Bottle for culture medium

          Ø  – maximum capacity 30 ml

          • 1 mounting tray to take 2 chambers

           

           

           

          Kirkstall Ltd, based in Sheffield, has just launched a new cell-culture system called Quasi-Vivo® that enables longer term in-vitro cell-culture.
          A research paper, to be published in the Journal of Hepatology, describes how primary human Hepatocytes maintained correct phenotype and metabolic activity over more than 28 days in the system. This breakthrough enabled the cells to grow in a standard medium without the need for a complex cocktail of growth factors. It follows a collaborative programme of research and evaluation of the technology with 5 leading centres for cellculture across Europe.
          The key to success is the flow system and the design of the cell culture chamber that together provide just the right level of nutrient and oxygen without causing flow stress.

           

           

          “This system removes the hassle that has plagued microfluidics and Lab-on-a-chip in-vitro culture for years,” said Dr Malcolm Wilkinson, CEO of Kirkstall. Cells can be seeded and cultured on cover slips or scaffolds inside the Quasi-Vivo® chambers.

          Interest in the technology is high. A recent meeting in Montpellier organised by Kirkstall brought together 80 researchers from Europe. Next year’s meeting will take place in May at Saarbrucken, hosted by Professor Claus-Michael Lehr, Head of the Helmholtz Institute for Pharmaceutical Research at Saarland University. The conference will cover applications in cell-culture, stem cell research, disease modelling drug discovery and regenerative medicine.

           

           

           

           

           

           

           

           

           

           

           

           

           

           

           

           

           

           

           

           

           

           

           

           

          The Kirkstall Quasi-Vivo® system enables multiple cell types to be cultured in inter-connected culture chambers. The nutrient flow between chambers makes it possible to mimic different metabolic pathways to investigate and test multi-compartmental biological models in vitro. We are looking for partners to help us develop better models of in vitro cell culture for studies of drug and chemical toxicity, disease, DMPK and stem cell differentiation.

          Deadline:

          10/29/2013

          Name of organisation:

          Kirkstall Ltd

          Executive Summary:

          Quasi-Vivo Cell Culture

          Kirkstall has recently launched the Quasi-Vivo® 500 Starter Kit for use by the research community. The dynamic nutrient flow means that unlike conventional static cell culture, cell to cell signalling is permitted between different cell and tissue types, whilst the bio-module design minimises the shear stress in the cell culture region, improving cell vitality and allowing longer duration experiments.

          Scientific background:

          Quasi-Vivo Cell Culture

          Traditionally, static cell culture systems using well plates or Petri dishes have been used to represent human or animal physiology. However, there is growing consensus that such 2D models are poor and inadequate predictors of in vivo responses (1, 2). Cells are sensitive to their microenvironment, which is rich in 3D cues from the extracellular matrix, other cells and from mechanical/chemical stimuli due to flow, concentration gradients and movement. Traditional methods for investigating cellular responses in vitro are inadequate in this sense, since the complex interplay of mechanical and biochemical factors is largely absent.

          When using 3D culture it is important to provide perfusion of nutrients and oxygen to sustain the cells and maintain the correct metabolic competency. The Quasi-Vivo® approach achieves this by providing flow in a leak proof system, which, because of the modular construction of chambers allows multi-tissue models to be interconnected, sharing nutrient media and hence facilitating cell to cell signalling. Cells can be cultured on a wide variety of scaffolds or even coated coverslips.

          The system allows cell biologists who are familiar with the use of multi-well plates for static cell culture  to design experiments that are more physiologically meaningful. Peer reviewed papers are now being published confirming the benefits of 3D cell culture under flow in the Quasi-Vivo®system (3).The metabolic competency is enhanced offering a route to more complex models of disease and epigenetics that match clinical results (2). Tests using the Quasi-Vivo® system have already demonstrated better correlation to clinical trial results than previous in vitro cell culture methods - results published as a poster at Rome AAT meeting 2009.

          Current state of the art:

          A large number of bioreactor systems for cell culture have been designed and described (4 - 9). In most cases, the bioreactors described are custom designed for specific requirements and necessitate the use of particular seeding methods or scaffolds with narrow dimensional and design specifications, limiting their use by the wider research community. They can range in size from large-scale bioartificial livers with several billions of cells and several millilitres of fluid down to microfluidic systems which enable the culture of different cell types in a shear stress controlled environment with a few hundred microliters of medium (10). In a micro bioreactor, the cell culture surface is generally around 0.5 – 0.8mm2 (11) and this tiny surface is seeded with a few thousand cells. Such a small number of cells, organized on a tiny surface can be only a rough approximation of an organ and cannot meaningfully predict in vivo physiology or pathophysiology (12). Another problem of micro bioreactors is that the percentage of area close to the edge of the system is higher than in a mm-sized surface. A large fraction of the cell population is found in a peripheral zone of the system where they have higher cytoskeletal tensions (13), and they may also have different viability or activity (14). So there is a need for systems which overcome these issues and provide a general purpose in vitro cell culture system for a wide variety of cell types at a scale that allows accurate modelling of human clinical behaviour either in response to drugs or disease modelling.

          Hepatocytes are the main orchestrators of metabolism and represent the starting point for most organ models, however in previous bioreactors, primary hepatocytes have proved difficult to maintain in vitro as they lose their ability to metabolise drugs due to down regulation of the genes (notably CYPs) responsible for phase I metabolism after three days. Vinci et al (3) evaluated the effect of medium flow rate over 21 days (up to 500 uL/min) on 32 different genes. Using the Quasi-Vivo® system they found that mRNA expression of genes involved in xenobiotic/drug metabolism and transport in 2-week-old cultures reached levels close to or higher than those measured in freshly isolated hepatocytes demonstrating the importance of flow in maintaining cell cultures in anin vivo-like state.

          What could your solution:
          be used for?

          The Quasi-Vivo® design principles are based on the allometric scaling of cell numbers and the mean residence times of molecules in metabolic tissues, as well as consideration of oxygen tension and shear stress, which together can be combined to establish organ and system models. By connecting together different chambers in series or parallel, it is possible to mimic different metabolic pathways and test multi-compartmental biological models in vitro without having to design dedicated equipment or culture chambers (15).

          Combined with allometric scaling, the Quasi-Vivo® system could also find applications in the following metabolic models:

          ·         biotransformation (hepatocytes)

          ·         gas exchange and biotransformation (hepatocytes, lung, epithelial cells)

          ·         absorption and biotransformation (hepatocytes, skin epithelia)

          ·         nutrient absorption and biotransformation (hepatocytes, intestinal epithelial cells)

          ·         biotransformation and nutrient transport (hepatocytes, endothelial cells)

          ·         biotransformation, nutrient transport and nutrient absorption (hepatocytes, endothelial cells, intestinal epithelial cells).

          For more information visit: http://kirkstall.org/index.php/quasi-vivo-system/benefits-of-kirkstalls-quasi-vivotm-system/.

          Need for collaboration:

          Kirkstall is providing a basic research tool for in vitro cell culture. We need partners who wish to develop better models of in vitro cell culture for studies of drug and chemical toxicity, disease, DMPK and stem cell differentiation. Research institutes, universities or companies from the pharmaceutical and chemical sectors with existing models of kidney, lung, heart, blood-brain barrier, liver, etc are invited to contact us to participate in collaborative research.

          Of particular interest are: (i) models for cell to cell and organ to organ interaction e.g. for recapitulating drug absorption and metabolism a linked model of lung/GI tract with liver and then target organ or for modelling indirect multi-organ toxicity a model with liver and target organ, and (ii) studies that require long term cell culture over 28 days. We would particularly like to work with groups who have established excellent single tissue or organ models and now wish to extend these to include organ to organ interactions.

          3Rs impact assessment:

          ·         Use of the Quasi-Vivo® system for testing of hepatotoxicity, multi-organ toxicity and drug metabolism in pharmaceutical development will reduce the need for in vivostudies, particularly on candidate drugs destined to fail later in development. Use of the system for toxicity testing also has broader potential in the chemicals and consumer products industry.

          ·         As experience and validation with known compounds and toxicities grows, the potential for the eventual replacement of animal models in some applications exists, for example rodents in early stage drug toxicity screening.

          ·         Human primary cells that more accuray represent the in vivo situation using the Quasi-Vivo® system could potentially replace the use of animals in later stage pre-clinical trials

          ·         Different cell types from a single animal donor can be used in multiple experiments, thereby reducing the number of animals sacrificed per experiment, for instance liver, kidney, heart and lung from one animal for four separate experiments on disease or toxicity.

           

          The applications cover the need for better tools for prediction of toxicity of drugs using human primary cells or cell lines prior to human clinical trials as part of the drug discovery and development process. Initially it is likely that these tests would have to be run in parallel with animal tests which are still mandatory under current legislation. However, in the longer term, as confidence in the new techniques grows and the scientific data becomes compelling, it is hoped that regulations can be changed. This could then have a major impact on the number of animals used once these new in vitro tests are accepted as a way of avoiding the need for animal studies.

           

          REFERENCES

          (1) Zhang S. Beyond the petri dish. Nat Biotechnol 2004; 22(2):151-152,

          (2)  Kirkpatrick J, Fuchs S, Hermanns I et al. Cell culture models of higher complexity in tissue engineering and regenerative medicine, Biomaterials 2007; 28(34):5193-5198

          (3) Bruna Vinci, Cédric Duret, Sylvie Klieber, Sabine Gerbal-Chaloin, Antonio Sa-Cunha, Sylvain Laporte, Bertrand Suc, Patrick Maurel, Arti Ahluwalia and Martine Daujat-Chavanieu. Modular bioreactor for primary human hepatocyte culture: Medium flow stimulates expression and activity of detoxification genes, Biotechnol. J. 2011, 6, 554–564

          (4) Dumont K, Yperman J, Verbeken E, Segers P, Meuris B.. Design of a new pulsatile bioreactor for tissue engineered aortic heart valve formation. Artif Organs 2002, 26:710–714

          (5) Fu Q, Wu C, Shen Y, Zheng S, Chen R, Effect of LIMK2 RNAi on reorganisation of actin cytoskeleton in osteoblasts induced by fluid sheer stress. J Biomech 2008; 41(5): 3225 – 3228

          (6) Martin I, Wendt D, Heberer M.. The role of bioreactors in tissue engineering. Trends Biotechnol 2004; 22:80–86

          (7) Miyakawa A, Dallan LAO, Lacchini S, Borin TF, Krieger JE.. Human saphenous vein organ culture under controlled hemodynamic conditions. Clinics 2008; 63(5):683–688

          (8) Morelli S, Salerno S, Rende M, Lopez LC, Favia P, Procino A, Memoli B, Andreucci VE, d’Agostino R, Drioli E, De Bartolo L. Human hepatocyte functions in galactosylated membrane bioreactor, J Membr Sci 2007; 302: 27 – 35

          (9) Powers MJ, Domandsky K, Kaazempur-Mofrad MR, Kalezi A, Capitano A, Upadhyaya A, Kurzawski P, Wack KE, Stolz DB, Kamm R, Griffith LG. A microfabricated array bioreactor for perfused 3D liver culture. Biotechnol Bioeng 2002a; 78:257–269

          (10) De Bartolo L, Jarosch-Von Schweder G, Haverich A, Bader A. A novel full-scale flat membrane bioreactor utilizing porcine hepatocytes: Cell viability and tissue-specific functions. Biotechnol Prog 2000; 16(1):102–108

          (11) Baudoin R, Corlu A, Griscom L, Legallais CE. Trends in the development of microfluidic cell biochips for in vitro hepatotoxicity. Toxicol In Vitro 2007; 21:535–544

          (12) Tingley SK.. High-throughput cell culture: A real-world evaluation. Innovat Pharm Technol 2006; February:54–58

          (13) McBeath R, Pirone DM, Nelson CM, Bhadriraju K, Chen CS. Cell shape, cytoskeletal tension, and RhoA regulate stem cell lineage commitment. Dev Cell 2004, 6(4):483–495

          (14) Francis K, Palsson BO. Effective intercellular communication distances are determined by the relative time constants for cyto/chemokine secretion and diffusion. Proc Natl Acad Sci USA 1997; 94:12258–12262

          (15) Mazzei D, Guzzardi MA, Giusti S, Ahluwalia A. A Low shear stress modular bioreactor for connected cell culture under high flow rates. Biotechnol Bioeng. 2010;106(1):127-37

           

           

          Keywords:

          Toxicity testing, multi-compartment bioreactor, disease modelling, tissue engineering

           

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