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4 results on '"Razan, Florence"'

3. Improvement of HepG2/C3a cell functions in a microfluidic biochip

Author

Jean-Matthieu Prot, Anne Corlu, Caroline Aninat, Céline Brochot, Eric Leclerc, Cécile Legallais, Laurent Griscom, Florence Razan, Christiane Guillouzo, Bio-MIcroSystèmes et BioSensors (SATIE-BIOMIS), Systèmes d'Information et d'Analyse Multi-Echelles (SIAME), Systèmes et Applications des Technologies de l'Information et de l'Energie (SATIE), École normale supérieure - Cachan (ENS Cachan)-Université Paris-Sud - Paris 11 (UP11)-Institut Français des Sciences et Technologies des Transports, de l'Aménagement et des Réseaux (IFSTTAR)-École normale supérieure - Rennes (ENS Rennes)-Université de Cergy Pontoise (UCP), Université Paris-Seine-Université Paris-Seine-Conservatoire National des Arts et Métiers [CNAM] (CNAM)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Cachan (ENS Cachan)-Université Paris-Sud - Paris 11 (UP11)-Institut Français des Sciences et Technologies des Transports, de l'Aménagement et des Réseaux (IFSTTAR)-École normale supérieure - Rennes (ENS Rennes)-Université de Cergy Pontoise (UCP), Université Paris-Seine-Université Paris-Seine-Conservatoire National des Arts et Métiers [CNAM] (CNAM)-Centre National de la Recherche Scientifique (CNRS)-Systèmes et Applications des Technologies de l'Information et de l'Energie (SATIE), Université Paris-Seine-Université Paris-Seine-Conservatoire National des Arts et Métiers [CNAM] (CNAM)-Centre National de la Recherche Scientifique (CNRS), Razan, Florence, Biomécanique et génie biomédical (BIM), Centre National de la Recherche Scientifique (CNRS), Foie, métabolismes et cancer, Université de Rennes (UR)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Université Paris-Seine-Université Paris-Seine-Conservatoire National des Arts et Métiers [CNAM] (CNAM), HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM)-HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM)-Centre National de la Recherche Scientifique (CNRS), Institut National de l'Environnement Industriel et des Risques (INERIS), Université de Rennes 1 (UR1), and Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )

Subjects
MESH: Cell Death, MESH: Anemia, Glutamine, Cell, Microfluidics, 02 engineering and technology, Applied Microbiology and Biotechnology, MESH: Tissue Engineering, MESH: Recombinant Proteins, MESH: Hepatocytes, MESH: Microfluidics, MESH: Genotype, chemistry.chemical_compound, Tissue engineering, PDMS, MESH: Dimethylpolysiloxanes, MESH: Double-Blind Method, MESH: Hepacivirus, Biochip, ComputingMilieux_MISCELLANEOUS, MESH: Treatment Outcome, 0303 health sciences, MESH: Middle Aged, Cell Death, 021001 nanoscience & nanotechnology, MESH: Hepatitis C, Chronic, MESH: Glucose, medicine.anatomical_structure, Biochemistry, MESH: Cell Survival, MESH: RNA, Viral, MESH: Nylons, MESH: Interferon-alpha, 0210 nano-technology, MESH: Viral Load, Biotechnology, microfluidic biochip, MESH: Antiviral Agents, MESH: Serine Proteinase Inhibitors, Cell Survival, [SPI.NANO] Engineering Sciences [physics]/Micro and nanotechnologies/Microelectronics, Bioengineering, Biology, liver, Cell Line, 03 medical and health sciences, MESH: Gene Expression Profiling, MESH: Ribavirin, Albumins, medicine, Humans, Dimethylpolysiloxanes, [SPI.NANO]Engineering Sciences [physics]/Micro and nanotechnologies/Microelectronics, 030304 developmental biology, MESH: Glutamine, MESH: Humans, MESH: Proline, Tissue Engineering, Gene Expression Profiling, MESH: Adult, [SDV.MHEP.HEG]Life Sciences [q-bio]/Human health and pathology/Hépatology and Gastroenterology, transcriptomic, Molecular biology, Fold change, In vitro, MESH: Male, MESH: Cell Line, MESH: Drug Therapy, Combination, Nylons, Glucose, chemistry, MESH: Polyethylene Glycols, Cell culture, Hepatocytes, MESH: African Continental Ancestry Group, MESH: Albumins, Xenobiotic, MESH: Female, Drug metabolism
Abstract

International audience; Current developments in tissue engineering and microtechnology fields allow the use of microfluidic biochip as microtools for in vitro investigations. In the present study, we describe the behavior of HepG2/C3a cells cultivated in a poly(dimethylsiloxane) (PDMS) microfluidic biochip coupled to a perfusion system. Cell culture in the microfluidic biochip for 96 h including 72 h of perfusion provoked a 24 h delay in cell growth compared to plate cultures. Inside the microfluidic biochip, few apoptosis, and necrosis were detected along the culture and 3D cell organization was observed. Regarding the hepatic metabolism, glucose and glutamine consumptions as well as albumin synthesis were maintained. A transcriptomic analysis performed at 96 h of culture using Affymetrix GeneChip demonstrated that 1,025 genes with a fold change above 1.8 were statistically differentially expressed in the microfluidic biochip cultures compared to plate cultures. Among those genes, phase I enzymes involved in the xenobiotic's metabolism such as the cytochromes P450 (CYP) 1A1/2, 2B6, 3A4, 3A5, and 3A7 were up-regulated. The CYP1A1/2 up-regulation was associated with the appearance of CYP1A1/2's activity evidenced by using EROD biotransformation assay. Several phase II enzymes such as sulfotransferases (SULT1A1 and SULT1A2), UDP-glucuronyltransferase (UGT1A1, UGT2B7) and phase III transporters (such as MDR1, MRP2) were also up-regulated. In conclusion, microfluidic biochip could and provide an important insight to exploring the xenobiotic's metabolism. Altogether, these results suggest that this kind of biochip could be considered as a new pertinent tool for predicting cell toxicity and clearance of xenobiotics in vitro.

Published
2010

4. Investigation of ifosfamide nephrotoxicity induced in a liver-kidney co-culture biochip.

Author

Choucha-Snouber L, Aninat C, Grsicom L, Madalinski G, Brochot C, Poleni PE, Razan F, Guillouzo CG, Legallais C, Corlu A, and Leclerc E

Subjects
Acetaldehyde analogs & derivatives, Acetaldehyde toxicity, Animals, Calcium metabolism, Cell Cycle drug effects, Cell Proliferation drug effects, Cell Shape drug effects, Dogs, Fluorescent Dyes, Gene Expression drug effects, Hep G2 Cells, Humans, Kidney cytology, Liver cytology, Madin Darby Canine Kidney Cells, Real-Time Polymerase Chain Reaction, Coculture Techniques methods, Ifosfamide toxicity, Kidney drug effects, Liver drug effects, Microfluidic Analytical Techniques methods, Tissue Array Analysis methods
Abstract

In this article, we present a liver-kidney co-culture model in a micro fluidic biochip. The liver was modeled using HepG2/C3a and HepaRG cell lines and the kidney using MDCK cell lines. To demonstrate the synergic interaction between both organs, we investigated the effect of ifosfamide, an anticancerous drug. Ifosfamide is a prodrug which is metabolized by the liver to isophosforamide mustard, an active metabolite. This metabolism process also leads to the formation of chloroacetaldehyde, a nephrotoxic metabolite and acrolein a urotoxic one. In the biochips of MDCK cultures, we did not detect any nephrotoxic effects after 72 h of 50 µM ifosfamide exposure. However, in the liver-kidney biochips, the same 72 h exposure leads to a nephrotoxicity illustrated by a reduction of the number of MDCK cells (up to 30% in the HepaRG-MDCK) when compared to untreated co-cultures or treated MDCK monocultures. The reduction of the MDCK cell number was not related to a modification of the cell cycle repartition in ifosfamide treated cases when compared to controls. The ifosfamide biotransformation into 3-dechloroethylifosfamide, an equimolar byproduct of the chloroacetaldehyde production, was detected by mass spectrometry at a rate of apparition of 0.3 ± 0.1 and 1.1 ± 0.3 pg/h/biochips in HepaRG monocultures and HepaRG-MDCK co-cultures respectively. Any metabolite was detected in HepG2/C3a cultures. Furthermore, the ifosfamide treatment in HepaRG-MDCK co-culture system triggered an increase in the intracellular calcium release in MDCK cells on contrary to the treatment on MDCK monocultures. As 3-dechloroethylifosfamide is not toxic, we have tested the effect of equimolar choloroacetaldehyde concentration onto the MDCK cells. At this concentration, we found a quite similar calcium perturbation and MDCK nephrotoxicity via a reduction of 30% of final cell numbers such as in the ifosfamide HepaRG-MDCK co-culture experiments. Our results suggest that ifosfamide nephrotoxicity in a liver-kidney micro fluidic co-culture model using HepaRG-MDCK cells is induced by the metabolism of ifosfamide into chloroacetaldehyde whereas this pathway is not functional in HepG2/C3a-MDCK model. This study demonstrates the interest in the development of systemic organ-organ interactions using micro fluidic biochips. It also illustrated their potential in future predictive toxicity model using in vitro models as alternative methods., (Copyright © 2012 Wiley Periodicals, Inc.)

Published
2013
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