title: HepaRG culture in tethered spheroids as an in vitro three-dimensional model for drug safety screening.
abstract: Conventional two-dimensional (2D) monolayer cultures of HepaRG cells allow in vitro maintenance of many liver-specific functions. However, cellular dedifferentiation and functional deterioration over an extended culture period in the conventional 2D HepaRG culture have hampered its applications in drug testing. To address this issue, we developed tethered spheroids of HepaRG cells on Arg-Gly-Asp (RGD) and galactose-conjugated substratum with an optimized hybrid ratio as an in vitro three-dimensional (3D) human hepatocyte model. The liver-specific gene expression level and drug metabolizing enzyme activities in HepaRG-tethered spheorids were markedly higher than those in 2D cultures throughout the culture period of 7 days. The inducibility of three major cytochrome P450 (CYP) enzymes, namely CYP1A2, CYP2B6 and CYP3A4, was improved in both mRNA and activity level in tethered spheroids. Drug-induced cytotoxic responses to model hepatotoxins (acetaminophen, chlorpromazine and ketoconazole) in tethered spheroids were comparable to 2D cultures as well as other studies in the literature. Our results suggested that the HepaRG-tethered spheroid would be an alternative in vitro model suitable for drug safety screening.
authors.affiliation: Department of Gastroenterology, Nanfang Hospital, Southern Medical University, No. 1838, North of Guangzhou Dadao, Guangzhou, 510515, China.|Department of Gastroenterology, Beijing Chao-Yang Hospital, Capital Medical University, No. 8, South Road of Workers' Gymnasium, Beijing, 100020, China.
authors.forename: Zenan
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Department of Gastroenterology, Nanfang Hospital, Southern Medical University, No. 1838, North of Guangzhou Dadao, Guangzhou, 510515, China.
authors.forename: Xiaobei
authors.initials: X
authors.lastname: Luo
authors.affiliation: Department of Bioengineering, National University of Singapore, Block EA, #03-12, 9 Engineering Drive 1, Singapore, 117576, Singapore.
authors.forename: Chukwuemeka
authors.initials: C
authors.lastname: Anene-Nzelu
authors.affiliation: Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, 10 Medical Drive, Clinical Research Center, #04-25, Singapore, 117597, Singapore.
authors.forename: Yu
authors.initials: Y
authors.lastname: Yu
authors.affiliation: Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, 10 Medical Drive, Clinical Research Center, #04-25, Singapore, 117597, Singapore.
authors.forename: Xin
authors.initials: X
authors.lastname: Hong
authors.affiliation: Institute of Bioengineering and Nanotechnology, A*STAR, The Nanos, #04-01, 31 Biopolis Way, Singapore, 138669, Singapore.
authors.forename: Nisha Hari
authors.initials: NH
authors.lastname: Singh
authors.affiliation: Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, 10 Medical Drive, Clinical Research Center, #04-25, Singapore, 117597, Singapore.
authors.forename: Lei
authors.initials: L
authors.lastname: Xia
authors.affiliation: Department of Gastroenterology, Nanfang Hospital, Southern Medical University, No. 1838, North of Guangzhou Dadao, Guangzhou, 510515, China.
authors.forename: Side
authors.initials: S
authors.lastname: Liu
authors.affiliation: Department of Bioengineering, National University of Singapore, Block EA, #03-12, 9 Engineering Drive 1, Singapore, 117576, Singapore.|Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, 10 Medical Drive, Clinical Research Center, #04-25, Singapore, 117597, Singapore.|Institute of Bioengineering and Nanotechnology, A*STAR, The Nanos, #04-01, 31 Biopolis Way, Singapore, 138669, Singapore.|NUS Graduate School for Integrative Sciences and Engineering, Centre for Life Sciences (CeLS), #05-01, 28 Medical Drive, Singapore, 117576, Singapore.|Singapore-MIT Alliance, Computational and System Biology Program, E4-04-10, 4 Engineering Drive 3, Singapore, 117576, Singapore.|NUS Tissue Engineering Program, DSO Labs, National University of Singapore, Singapore, 117597, Singapore.|Singapore-MIT Alliance for Research and Technology, 3 Science Drive 2, S16-05-08, Singapore, 117543, Singapore.|Department of Biological Engineering, Massachusetts Institute of Technology, 77 Massachusetts Ave., Cambridge, MA, 02139, USA.
authors.forename: Hanry
authors.initials: H
authors.lastname: Yu
mesh_terms: D018929:Cell Culture Techniques; D002470:Cell Survival / Q000187:drug effects; D056486:Chemical and Drug Induced Liver Injury / Q000473:pathology; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis / Q000235:genetics; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D004790:Enzyme Induction / Q000187:drug effects; D005690:Galactose / Q000378:metabolism; D022781:Hepatocytes / Q000187:drug effects; D006801:Humans; D008954:Models, Biological; D009842:Oligopeptides / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis; D018874:Spheroids, Cellular / Q000187:drug effects* / Q000648:ultrastructure; D018675:Toxicity Tests / Q000379:methods*; D014407:Tumor Cells, Cultured
chemical_list: D009842:Oligopeptides; D012333:RNA, Messenger; C047981:arginyl-glycyl-aspartic acid; D003577:Cytochrome P-450 Enzyme System; D005690:Galactose
keywords: 3D in vitro model; CYP induction; HepaRG; RGD; drug safety testing; galactose; xenobiotics