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Article ; Online: Expression and functional activity of cytochrome P450 enzymes in human hepatocytes with sustainable reproducibility for in vitro phenotyping studies.

Bachour-El Azzi, Pamela / Chesné, Christophe / Uehara, Shotaro

Advances in pharmacology (San Diego, Calif.)

2022 Volume 95, Page(s) 285–305

Abstract: Primary human hepatocytes are an essential in vitro tool for evaluating drug metabolism, drug-drug interactions, and hepatotoxicity. This model is considered as the gold standard in matter of DMPK studies in both industrial and academic research. The ... ...

Abstract	Primary human hepatocytes are an essential in vitro tool for evaluating drug metabolism, drug-drug interactions, and hepatotoxicity. This model is considered as the gold standard in matter of DMPK studies in both industrial and academic research. The primary human hepatocytes are used either in suspension or in monolayer, as fresh or frozen cells. However, the use of this model is limited due to the lack of availability, rapid loss of functionality, high cost as well as the variable hepatocyte plating efficiencies in culture and the limited stock of hepatocytes derived from the same origin. Chimeric TK-NOG mice with humanized livers (humanized liver mice) are an attractive platform for drug metabolism and toxicity, which were produced by transplanting human hepatocytes into immunodeficient mice with injured livers. Here, we show that, using humanized mouse liver, in vivo human hepatocyte repopulation was over ~100-fold enabling the continuous and abundant use of human hepatocytes of the same origin and improving their plateability. In our latest cell preparations, hepatocytes isolated from humanized liver mice (Hu-Liver cells) exhibited high purity (ratio of HLA-positive cells: 92±3%), good viability (75±12%), and yield (1.0×10
MeSH term(s)	Animals ; Cells, Cultured ; Cytochrome P-450 Enzyme System/genetics ; Cytochrome P-450 Enzyme System/metabolism ; Hepatocytes ; Humans ; Liver/metabolism ; Mice ; Reproducibility of Results
Chemical Substances	Cytochrome P-450 Enzyme System (9035-51-2)
Language	English
Publishing date	2022-06-30
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1557-8925
ISSN (online)	1557-8925
DOI	10.1016/bs.apha.2022.05.009
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Article ; Online: Contribution of Humanized Liver Chimeric Mice to the Study of Human Hepatic Drug Transporters: State of the Art and Perspectives.

Zerdoug, Anna / Le Vée, Marc / Uehara, Shotaro / Lopez, Béatrice / Chesné, Christophe / Suemizu, Hiroshi / Fardel, Olivier

European journal of drug metabolism and pharmacokinetics

2022 Volume 47, Issue 5, Page(s) 621–637

Abstract: Chimeric mice with humanized livers constitute an attractive emergent experimental model for investigating human metabolism and disposition of drugs. The present review was designed to summarize key findings about the use of this model for studying human ...

Abstract	Chimeric mice with humanized livers constitute an attractive emergent experimental model for investigating human metabolism and disposition of drugs. The present review was designed to summarize key findings about the use of this model for studying human hepatic drug transporters, which are now recognized as important players in pharmacokinetics and consequently have to be considered from a regulatory perspective during pharmaceutical drug development. The reviewed data indicate that chimeric mice with humanized livers have been successfully used for analysing the implications of human hepatic drug transporters for drug hepatobiliary elimination, drug-drug interactions and drug-induced cholestasis. Such transporter studies have been performed in vivo with chimeric mice and/or in vitro with human hepatocytes isolated from humanized liver and used either in suspension or in culture. The residual presence of mouse hepatocytes and the potential morphological/histological alterations of the humanized liver, as well as its immunodeficient mouse environment, have, however, to be considered when using chimeric mice with humanized livers for transporter studies. Finally, if the proof of concept of applying chimeric mice with humanized livers to hepatic drug transport is established, more experimental data on this topic, including from standardization approaches, are likely required to completely and accurately demonstrate the robustness, convenience and added value of this chimeric mouse model for drug transporter studies.
MeSH term(s)	Animals ; Chimera/metabolism ; Hepatocytes/metabolism ; Humans ; Liver/metabolism ; Membrane Transport Proteins/metabolism ; Metabolic Clearance Rate ; Mice
Chemical Substances	Membrane Transport Proteins
Language	English
Publishing date	2022-07-06
Publishing country	France
Document type	Journal Article ; Review
ZDB-ID	196729-0
ISSN	2107-0180 ; 0398-7639 ; 0378-7966
ISSN (online)	2107-0180
ISSN	0398-7639 ; 0378-7966
DOI	10.1007/s13318-022-00782-9
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Article: How to Foster ‘New Approach Methodology’ Toxicologists

Doktorova, Tatyana Y. / Azzi, Pamela / Hofer, Joelle / Messner, Catherine J. / Gaiser, Carine / Werner, Sophie / Singh, Pranika / Hardy, Barry / Suter-Dick, Laura / Chesne, Christophe

ATLA. Alternatives to laboratory animals. , v. 50, no. 1

2022

Abstract: The need to reduce, refine and replace animal experimentation has led to a boom in the establishment of new approach methodologies (NAMs). This promising trend brings the hope that the replacement of animals by using NAMs will become increasingly ... ...

Abstract	The need to reduce, refine and replace animal experimentation has led to a boom in the establishment of new approach methodologies (NAMs). This promising trend brings the hope that the replacement of animals by using NAMs will become increasingly accepted by regulators, included in legislation, and consequently more-often implemented by industry. The majority of NAMs, however, are still not very well understood, either due to the complexity of the applied approach or the data analysis workflow. A potential solution to this problem is the provision of better educational resources to scientists new to the area — showcasing the added value of NAMs and outlining various ways of overcoming issues associated with knowledge gaps. In this paper, the educational exchange between four institutions — namely, two universities and two SMEs — via a series of video training sessions, is described. The goal of this exchange was to showcase an exemplary event to help introduce scientists to non-animal approaches, and to actively support the development of resources enabling the use of alternatives to laboratory animals.
Keywords	animal experimentation ; industry ; laboratories ; laws and regulations
Language	English
Dates of publication	2022-01
Size	p. 71-75.
Publishing place	SAGE Publications
Document type	Article
ZDB-ID	605800-0
ISSN	0261-1929
ISSN	0261-1929
DOI	10.1177/02611929221078945
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Article ; Online: Drug transporter expression and activity in cryopreserved human hepatocytes isolated from chimeric TK-NOG mice with humanized livers.

Zerdoug, Anna / Le Vée, Marc / Uehara, Shotaro / Jamin, Agnès / Higuchi, Yuichiro / Yoneda, Nao / Lopez, Béatrice / Chesné, Christophe / Suemizu, Hiroshi / Fardel, Olivier

Toxicology in vitro : an international journal published in association with BIBRA

2023 Volume 90, Page(s) 105592

Abstract: Chimeric mice with humanized liver are thought to represent a sustainable source of isolated human hepatocytes for in vitro studying detoxification of drugs in humans. Because drug transporters are now recognized as key-actors of the hepatic detoxifying ... ...

Abstract	Chimeric mice with humanized liver are thought to represent a sustainable source of isolated human hepatocytes for in vitro studying detoxification of drugs in humans. Because drug transporters are now recognized as key-actors of the hepatic detoxifying process, the present study was designed to characterize mRNA expression and activity of main hepatic drug transporters in cryopreserved human hepatocytes isolated from chimeric TK-NOG mice and termed HepaSH cells. Such cells after thawing were shown to exhibit a profile of hepatic solute carrier (SLC) and ATP-binding cassette (ABC) drug transporter mRNA levels well correlated to those found in cryopreserved primary human hepatocytes or human livers. HepaSH cells used either as suspensions or as 24 h-cultures additionally displayed notable activities of uptake SLCs, including organic anion transporting polypeptides (OATPs), organic anion transporter 2 (OAT2) or sodium-taurocholate co-transporting polypeptide (NTCP). SLC transporter mRNA expression, as well as SLC activities, nevertheless fell in HepaSH cells cultured for 120 h, which may reflect a partial dedifferentiation of these cells with time in culture in the conventional monolayer culture conditions used in the study. These data therefore support the use of cryopreserved HepaSH cells as either suspensions or short-term cultures for drug transport studies.
MeSH term(s)	Humans ; Mice ; Animals ; Suspensions ; Liver/metabolism ; Hepatocytes/metabolism ; Membrane Transport Proteins/genetics ; Membrane Transport Proteins/metabolism ; Organic Anion Transporters/genetics ; Organic Anion Transporters/metabolism ; ATP-Binding Cassette Transporters/metabolism ; RNA, Messenger/metabolism
Chemical Substances	Suspensions ; Membrane Transport Proteins ; Organic Anion Transporters ; ATP-Binding Cassette Transporters ; RNA, Messenger
Language	English
Publishing date	2023-04-07
Publishing country	England
Document type	Journal Article
ZDB-ID	639064-x
ISSN	1879-3177 ; 0887-2333
ISSN (online)	1879-3177
ISSN	0887-2333
DOI	10.1016/j.tiv.2023.105592
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Zs.A 2223: Show issues

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Article ; Online: OECD workshop consensus report: Ethical considerations with respect to human derived products, specifically human serum, in OECD test guidelines.

Jacobs, Miriam N / Bult, Jan M / Cavanagh, Kevin / Chesne, Christophe / Delrue, Nathalie / Fu, Jianan / Grange, Emma / Langezaal, Ingrid / Misztela, Dominika / Murray, Jenny / Paparella, Martin / Stoddart, Gilly / Tonn, Torsten / Treasure, Carol / Tsukano, Masaaki / Versteegen, Rosemary

Frontiers in toxicology

2023 Volume 5, Page(s) 1140698

Abstract: The ethical needs and concerns with use and sourcing of human materials, particularly serum, in ... ...

Abstract	The ethical needs and concerns with use and sourcing of human materials, particularly serum, in OECD
Language	English
Publishing date	2023-02-27
Publishing country	Switzerland
Document type	Journal Article
ISSN	2673-3080
ISSN (online)	2673-3080
DOI	10.3389/ftox.2023.1140698
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Article ; Online: Liposome-Mediated Gene Transfer in Differentiated HepaRG™ Cells: Expression of Liver Specific Functions and Application to the Cytochrome P450 2D6 Expression.

Vlach, Manuel / Coppens-Exandier, Hugo / Jamin, Agnès / Berchel, Mathieu / Scaviner, Julien / Chesné, Christophe / Montier, Tristan / Jaffrès, Paul-Alain / Corlu, Anne / Loyer, Pascal

Cells

2022 Volume 11, Issue 23

Abstract: The goal of this study was to establish a procedure for gene delivery mediated by cationic liposomes in quiescent differentiated HepaRG™ human hepatoma cells. We first identified several cationic lipids promoting efficient gene transfer with low toxicity ...

Abstract	The goal of this study was to establish a procedure for gene delivery mediated by cationic liposomes in quiescent differentiated HepaRG™ human hepatoma cells. We first identified several cationic lipids promoting efficient gene transfer with low toxicity in actively dividing HepG2, HuH7, BC2 and progenitor HepaRG™ human hepatoma cells. The lipophosphoramidate Syn1-based nanovector, which allowed the highest transfection efficiencies of progenitor HepaRG™ cells, was next used to transfect differentiated HepaRG™ cells. Lipofection of these cells using Syn1-based liposome was poorly efficient most likely because the differentiated HepaRG™ cells are highly quiescent. Thus, we engineered the differentiated HepaRG™ Mitogenic medium supplement (ADD1001) that triggered robust proliferation of differentiated cells. Importantly, we characterized the phenotypical changes occurring during proliferation of differentiated HepaRG™ cells and demonstrated that mitogenic stimulation induced a partial and transient decrease in the expression levels of some liver specific functions followed by a fast recovery of the full differentiation status upon removal of the mitogens. Taking advantage of the proliferation of HepaRG™ cells, we defined lipofection conditions using Syn1-based liposomes allowing transient expression of the cytochrome P450 2D6, a phase I enzyme poorly expressed in HepaRG cells, which opens new means for drug metabolism studies in HepaRG™ cells.
Language	English
Publishing date	2022-12-02
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2661518-6
ISSN	2073-4409 ; 2073-4409
ISSN (online)	2073-4409
ISSN	2073-4409
DOI	10.3390/cells11233904
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Article ; Online: The Objective Stretch Marks Photonumeric Assessment Scale: A New and Complete Method to Assess Striae Distensae.

La Padula, Simone / Hersant, Barbara / Pizza, Chiara / Chesné, Christophe / Jamin, Agnes / Ben Mosbah, Ismail / D'Andrea, Francesco / Persichetti, Paolo / Rega, Umberto / Pensato, Rosita / Meningaud, Jean Paul

Plastic and reconstructive surgery

2022 Volume 151, Issue 2, Page(s) 307–313

Abstract: Background: Striae distensae evaluation criteria have been recently described, but none is focused on objective striae assessment. With the purpose of better and objectively estimating the severity of striae distensae, the Objective Stretch Marks ... ...

Abstract	Background: Striae distensae evaluation criteria have been recently described, but none is focused on objective striae assessment. With the purpose of better and objectively estimating the severity of striae distensae, the Objective Stretch Marks Assessment Scale has been developed by the authors' team. Methods: Seven hundred White patients were included in the study and assessed. To assess the severity of striae distensae, abdomen, breasts, hips, gluteal area, back area, thighs, calves, and upper limbs photonumeric grading scales were developed. The Rasch model was used as part of the validation process. A score was attributed to each patient, based on the scales we developed. The interrater reliability and test-retest reliability were analyzed. Results: Eight photonumeric scales for striae distensae treatment outcomes assessment were developed. All scales exceeded criteria for acceptability, reliability and validity. The interrater and intrarater reliabilities were good, with a substantial or virtually perfect interrater reliability for the total score (P = 0.16). Conclusions: The authors' results allowed them to validate the Objective Stretch Marks Assessment Scale as a reliable and reproducible tool to assess striae distensae treatment outcomes. This scale could be also considered as an important new metric that can be used in clinical research.
MeSH term(s)	Humans ; Striae Distensae/diagnosis ; Striae Distensae/therapy ; Reproducibility of Results ; Breast ; Treatment Outcome ; Abdomen
Language	English
Publishing date	2022-11-09
Publishing country	United States
Document type	Journal Article
ZDB-ID	208012-6
ISSN	1529-4242 ; 0032-1052 ; 0096-8501
ISSN (online)	1529-4242
ISSN	0032-1052 ; 0096-8501
DOI	10.1097/PRS.0000000000009835
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Uk I Zs.142: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 2021: Bestellungen von Artikeln über das Online-Bestellformular ab Jg. 2022: Lesesaal (EG)
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Article: How to Foster 'New Approach Methodology' Toxicologists.

Doktorova, Tatyana Y / Azzi, Pamela / Hofer, Joelle / Messner, Catherine J / Gaiser, Carine / Werner, Sophie / Singh, Pranika / Hardy, Barry / Suter-Dick, Laura / Chesne, Christophe

Alternatives to laboratory animals : ATLA

2022 Volume 50, Issue 1, Page(s) 71–75

Abstract	The need to reduce, refine and replace animal experimentation has led to a boom in the establishment of new approach methodologies (NAMs). This promising trend brings the hope that the replacement of animals by using NAMs will become increasingly accepted by regulators, included in legislation, and consequently more-often implemented by industry. The majority of NAMs, however, are still not very well understood, either due to the complexity of the applied approach or the data analysis workflow. A potential solution to this problem is the provision of better educational resources to scientists new to the area - showcasing the added value of NAMs and outlining various ways of overcoming issues associated with knowledge gaps. In this paper, the educational exchange between four institutions - namely, two universities and two SMEs - via a series of video training sessions, is described. The goal of this exchange was to showcase an exemplary event to help introduce scientists to non-animal approaches, and to actively support the development of resources enabling the use of alternatives to laboratory animals.
MeSH term(s)	Animal Experimentation ; Animal Testing Alternatives/methods ; Animals ; Universities
Language	English
Publishing date	2022-02-18
Publishing country	England
Document type	Journal Article
ZDB-ID	605800-0
ISSN	0261-1929
ISSN	0261-1929
DOI	10.1177/02611929221078945
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Article ; Online: Human HepaRG Cells can be Cultured in Hanging-drop Plates for Cytochrome P450 Induction and Function Assays.

Murayama, Norie / Usui, Takashi / Slawny, Nicky / Chesné, Christophe / Yamazaki, Hiroshi

Drug metabolism letters

2015 Volume 9, Issue 1, Page(s) 3–7

Abstract: Recent guidance/guidelines for industry recommend that cytochrome P450 induction can be assessed using human hepatocyte enzyme activity and/or mRNA levels to evaluate potential drug- drug interactions. To evaluate time-dependent cytochrome P450 induction ...

Abstract	Recent guidance/guidelines for industry recommend that cytochrome P450 induction can be assessed using human hepatocyte enzyme activity and/or mRNA levels to evaluate potential drug- drug interactions. To evaluate time-dependent cytochrome P450 induction precisely, induction of CYP1A2, CYP2B6, and CYP3A4 mRNA was confirmed (>2-fold) by the treatment with omeprazole, phenobarbital, and rifampicin, respectively, for 24 or 48 h on day 3 from the start of culture. After 24 h, the fold induction of CYP1A2 with 3.6 and 1.8x10(4) HepaRG cells per well was lower than that for 7.2x10(4) cells. CYP1A2 induction levels at 24 h were higher than those after 48 h. In contrast, higher CYP2B6 inductions were confirmed after 48 h exposure than after 24 h, independent of the number of cells per well. To help reduce the use of human cryopreserved hepatocytes, typical P450-dependent enzyme activities were investigated in human HepaRG cells cultured in commercial hanging-drop plates. Newly designed 96-well hanging-drop plates were capable of maintaining human CYP3A-dependent midazolam hydroxylation activities for up to 4 days using only 10% of the recommended initial 7.2x10(4) cells per well. Favorable HepaRG function using hanging-drop plates was confirmed by detecting 1'- hydroxymidazolam O-glucuronide on day 3, suggesting an improvement over traditional control plates in which this metabolite can be detected for 24-well plates. These results suggest that the catalytic function and/or induction of CYP1A2, CYP2B6, and CYP3A4 can be readily assessed with reduced numbers of starting HepaRG cells cultured in three-dimensional cultures in drops prepared with hanging-drop plates.
MeSH term(s)	Cell Culture Techniques ; Cytochrome P-450 CYP1A2/biosynthesis ; Cytochrome P-450 CYP2B6/biosynthesis ; Cytochrome P-450 CYP3A/biosynthesis ; Cytochrome P-450 Enzyme Inducers/pharmacology ; Cytochrome P-450 Enzyme System/biosynthesis ; Cytochrome P-450 Enzyme System/genetics ; Drug Interactions ; Enzyme Induction ; Hep G2 Cells ; Hepatocytes/drug effects ; Hepatocytes/enzymology ; Humans ; RNA, Messenger/biosynthesis ; Risk Assessment ; Risk Factors ; Substrate Specificity ; Time Factors
Chemical Substances	Cytochrome P-450 Enzyme Inducers ; RNA, Messenger ; Cytochrome P-450 Enzyme System (9035-51-2) ; CYP1A2 protein, human (EC 1.14.14.1) ; CYP2B6 protein, human (EC 1.14.14.1) ; Cytochrome P-450 CYP1A2 (EC 1.14.14.1) ; Cytochrome P-450 CYP2B6 (EC 1.14.14.1) ; Cytochrome P-450 CYP3A (EC 1.14.14.1) ; CYP3A4 protein, human (EC 1.14.14.55)
Language	English
Publishing date	2015-01-19
Publishing country	United Arab Emirates
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1874-0758
ISSN (online)	1874-0758
DOI	10.2174/1872312809666150119104806
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Article ; Online: Tunisian Clematis flammula Essential Oil Enhances Wound Healing: GC-MS Analysis, Biochemical and Histological Assessment.

Saidi, Rakia / Ghrab, Ferdaws / Kallel, Rim / Feki, Abdelfattah El / Boudawara, Tahya / Chesné, Christophe / Ammar, Emna / Jarraya, Raoudha Mezghani

Journal of oleo science

2019 Volume 67, Issue 11, Page(s) 1483–1499

Abstract: The aerial part of Clematis flammula (Ranunculaceae) has been traditionally used in the treatment of skin diseases including mycotic infection in the Tunisian traditional medicine. The study was undertaken to extract and determine the essential oil ... ...

Abstract	The aerial part of Clematis flammula (Ranunculaceae) has been traditionally used in the treatment of skin diseases including mycotic infection in the Tunisian traditional medicine. The study was undertaken to extract and determine the essential oil chemical composition of Clematis flammula aerial parts and to assess the potential of anemonin in wound healing on mechanically wounded wistar rats. The essential oil was obtained by hydrodistillation and analyzed by GC-MS. Anemonin was isolated and then incorporated as active in a cream for which the cytotoxicity was evaluated by methyl thiazolyl tetrazolium (MTT)-based colorimetric assay. Then, its potential in wound healing on mechanically wounded wistar rats was assessed. The GC-MS analysis showed that the major compound was protoanemonin (86.74%) which spontaneously dimerised in part to form the anemonin. The wound healing activity of anemonin cream exhibited a non toxic potential of anemonin at a concentration of 25 µg/mL with a cell migration efficiency that reaches more than 80% after 48 hours of treatment. Wound healing efficiency was evaluated by monitoring morphological and skin histological analyses. Comparable wound surface reduction of the group treated by anemonin cream (p ≥ 0.05) when compared to the reference treated group. The skin histological analysis showed the completely wound closure. Antioxidant activity was assessed by the malondialdehyde (MDA) rates and antioxidant enzymes (glutathione peroxidase (GPx) and catalase) determination. The results provided strong support for the effective wound healing activity of anemonin cream, making it a promising candidate as a therapeutic agent in tissue repairing processes.
MeSH term(s)	Administration, Topical ; Animals ; Antioxidants/metabolism ; Catalase/metabolism ; Clematis/chemistry ; Female ; Furans/administration & dosage ; Furans/isolation & purification ; Furans/pharmacology ; Gas Chromatography-Mass Spectrometry ; Glutathione Peroxidase/metabolism ; Malondialdehyde/metabolism ; Oils, Volatile/administration & dosage ; Oils, Volatile/isolation & purification ; Oils, Volatile/pharmacology ; Rats, Wistar ; Skin/metabolism ; Skin/pathology ; Skin Cream ; Stimulation, Chemical ; Tunisia ; Wound Healing/drug effects
Chemical Substances	Antioxidants ; Furans ; Oils, Volatile ; Malondialdehyde (4Y8F71G49Q) ; protoanemonin (66FQZ1A5SO) ; Catalase (EC 1.11.1.6) ; Glutathione Peroxidase (EC 1.11.1.9) ; anemonin (G99XG5B674)
Language	English
Publishing date	2019-01-18
Publishing country	Japan
Document type	Journal Article
ISSN	1347-3352
ISSN (online)	1347-3352
DOI	10.5650/jos.ess18056
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