title: Tethered spheroids as an in vitro hepatocyte model for drug safety screening.
abstract: Hepatocyte spheroids mimic many in vivo liver-tissue phenotypes but increase in size during extended culture which limits their application in drug testing applications. We have developed an improved hepatocyte 3D spheroid model, namely tethered spheroids, on RGD and galactose-conjugated membranes using an optimized hybrid ratio of the two bioactive ligands. Cells in the spheroid configuration maintained 3D morphology and uncompromised differentiated hepatocyte functions (urea and albumin production), while the spheroid bottom was firmly tethered to the substratum maintaining the spheroid size in multi-well plates. The oblate shape of the tethered spheroids, with an average height of 32 μm, ensured efficient nutrient, oxygen and drug access to all the cells within the spheroid structure. Cytochrome P450 induction by prototypical inducers was demonstrated in the tethered spheroids and was comparable or better than that observed with hepatocyte sandwich cultures. These data suggested that tethered 3D hepatocyte spheroids may be an excellent alternative to 2D hepatocyte culture models for drug safety applications.
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.forename: Rashidah Binte
authors.initials: RB
authors.lastname: Sakban
authors.forename: Yinghua
authors.initials: Y
authors.lastname: Qu
authors.forename: Xin
authors.initials: X
authors.lastname: Hong
authors.forename: Wenxia
authors.initials: W
authors.lastname: Zhang
authors.forename: Bramasta
authors.initials: B
authors.lastname: Nugraha
authors.forename: Wen Hao
authors.initials: WH
authors.lastname: Tong
authors.forename: Abhishek
authors.initials: A
authors.lastname: Ananthanarayanan
authors.forename: Baixue
authors.initials: B
authors.lastname: Zheng
authors.forename: Ian Yin-Yan
authors.initials: IY
authors.lastname: Chau
authors.forename: Ruirui
authors.initials: R
authors.lastname: Jia
authors.forename: Michael
authors.initials: M
authors.lastname: McMillian
authors.forename: Jose
authors.initials: J
authors.lastname: Silva
authors.forename: Shannon
authors.initials: S
authors.lastname: Dallas
authors.forename: Hanry
authors.initials: H
authors.lastname: Yu
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D022781:Hepatocytes / Q000166:cytology* / Q000502:physiology; D006801:Humans; D008297:Male; D008954:Models, Biological; D051381:Rats; D017208:Rats, Wistar; D018874:Spheroids, Cellular / Q000166:cytology / Q000502:physiology*
chemical_list: D003094:Collagen