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Human hepatocyte functions in a crossed hollow fiber membrane bioreactor
Authors:Loredana De Bartolo  Simona Salerno  Efrem Curcio  Antonella Piscioneri  Maria Rende  Sabrina Morelli  Franco Tasselli  Augustinus Bader  Enrico Drioli
Affiliation:1. Institute on Membrane Technology, National Research Council of Italy, ITM-CNR, c/o University of Calabria, Via P. Bucci, Cubo 17/C, I-87030 Rende (CS), Italy;2. Department of Chemical Engineering and Materials, University of Calabria, Via P. Bucci, I-87030 Rende (CS), Italy;3. Department of Cell Biology, University of Calabria, Via P. Bucci, 87030 Rende (CS), Italy;4. Biomedical-Biotechnological Center, BBZ, University of Leipzig, Germany;1. Section of Experimental Oncology and Nanomedicine (SEON), Else Kröner-Fresenius Stiftungsprofessur for Nanomedicine, University Hospital Erlangen, Erlangen, Germany;2. Chair of Magnetofluiddynamics, Measuring and Automation Technology, Technische Universität Dresden, Dresden, Germany;1. Section of Translational Medicine, Division of Applied Medicine, Polwarth Building, Foresterhill, Aberdeen AB25 2ZD, UK;2. Department of Biomedical Engineering, Bioengineering Unit, University of Strathclyde, Glasgow G4 0NW, UK;3. Biologie Servier, Drug Safety Research Centre, 905 Route de Saran, 45520 Gidy, France;1. Dept. of Environ. Eng., Inha University, Namgu, Inharo 100, Incheon, Republic of Korea;2. Samsung SDI, 56, Gosan-ro, Uiwang-si, Gyeonggi-do 437-711, Republic of Korea;3. Dept. of Civil and Environ. Eng., Stanford University, Stanford, CA 94305, USA;1. Department of Environmental and Chemical Engineering, University of Calabria, via P. Bucci, 87036 Rende (CS), Italy;2. Department of Mechanical Engineering, University of Perugia, Via Duranti 1, 06123 Perugia, Italy;3. Department of Medical and Surgical Sciences, Magna Graecia University, Viale Europa – Germaneto, 88100 Catanzaro, Italy;4. Casa di Cura Villa del Sole, Viale Pio X 202, 88100 Catanzaro, Italy;5. Department of Mechanical and Aerospatial Engineering, Politecnico di Torino, Corso Duca degli Abruzzi 24, 10129 Torino, Italy;1. Department of Computing, Imperial College London, South Kensington SW7 2AZ, UK;2. Department of Chemical Engineering, Imperial College London, South Kensington SW7 2AZ, UK;3. Department of Haematology, Imperial College London, Northwick Park and St. Mark''s Campus HA1 3UJ, UK;1. Surface Nanoscience Group, Department of Physics, University of Calabria, Via P. Bucci Cubo 31C, I-87036 Rende, Cosenza, Italy;2. UNICARIBE Research Center, University of Calabria, I-87036 Rende, Cosenza, Italy;3. INFN, Sezione LNF, Gruppo Collegato di Cosenza, Via P. Bucci, I-87036 Rende, Cosenza, Italy;4. Laboratorio de Nanotecnología, Área de Ciencias Básicas y Ambientales, Instituto Tecnológico de Santo Domingo, Av. Los Próceres, 10602 Santo Domingo, Dominican Republic;5. CompNano, School of Physical Sciences and Nanotechnology, Yachay Tech University, EC-100119 Urcuquí, Ecuador;6. Escuela de Ciencias Naturales y Exactas, Pontificia Universidad Católica Madre y Maestra, Autopista Duarte Km 1 ½, Santiago de los Caballeros 5100, Dominican Republic;7. Institute on Membrane Technology, National Research Council, Via P. Bucci Cubo 17C, I-87036 Rende, Cosenza, Italy;8. Department of Environmental Engineering, University of Calabria, zia P. Bucci Cubo 44A, I-87036 Rende, Cosenza, Italy
Abstract:An important challenge in liver tissue engineering is the development of bioartificial systems that are able to favour the liver reconstruction and to modulate liver cell behaviour.A crossed hollow fiber membrane bioreactor was developed to support the long-term maintenance and differentiation of human hepatocytes. The bioreactor consists of two types of hollow fiber (HF) membranes with different molecular weight cut-off (MWCO) and physico-chemical properties cross-assembled in alternating manner: modified polyetheretherketone (PEEK-WC) and polyethersulfone (PES), used for the medium inflow and outflow, respectively. The combination of these two fiber set produces an extracapillary network for the adhesion of cells and a high mass exchange through the cross-flow of culture medium. The transport of liver specific products such as albumin and urea together with the transport of drug such as diazepam was modelled and compared with the experimental metabolic data. The theoretical metabolite concentration differed 7.5% for albumin and 5% for urea with respect to experimental data. The optimised perfusion conditions of the bioreactor allowed the maintenance of liver functions in terms of urea synthesis, albumin secretion and diazepam biotransformation up to 18 days of culture. In particular the good performance of the bioreactor was confirmed by the high rate of urea synthesis (28.7 μg/h 106 cells) and diazepam biotransformation. In the bioreactor human hepatocytes expressed at high levels the individual cytochrome P450 isoenzymes involved in the diazepam metabolism. The results demonstrated that crossed HF membrane bioreactor is able to support the maintenance of primary human hepatocytes preserving their liver specific functions for all investigated period. This device may be a potential tool in the liver tissue engineering for drug metabolism/toxicity testing and study of disease pathogenesis alternatively to animal experimentation.
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