title: Galactosylated cellulosic sponge for multi-well drug safety testing.
abstract: Hepatocyte spheroids can maintain mature differentiated functions, but collide to form bulkier structures when in extended culture. When the spheroid diameter exceeds 200 μm, cells in the inner core experience hypoxia and limited access to nutrients and drugs. Here we report the development of a thin galactosylated cellulosic sponge to culture hepatocytes in multi-well plates as 3D spheroids, and constrain them within a macroporous scaffold network to maintain spheroid size and prevent detachment. The hydrogel-based soft sponge conjugated with galactose provided suitable mechanical and chemical cues to support rapid formation of hepatocyte spheroids with a mature hepatocyte phenotype. The spheroids tethered in the sponge showed excellent maintenance of 3D cell morphology, cell-cell interaction, polarity, metabolic and transporter function and/or expression. For example, cytochrome P450 (CYP1A2, CYP2B2 and CYP3A2) activities were significantly elevated in spheroids exposed to β-naphthoflavone, phenobarbital, or pregnenolone-16α-carbonitrile, respectively. The sponge also exhibits minimal drug absorption compared to other commercially available scaffolds. As the cell seeding and culture protocols are similar to various high-throughput 2D cell-based assays, this platform is readily scalable and provides an alternative to current hepatocyte platforms used in drug safety testing applications.
authors: NUS Graduate School for Integrative Sciences and Engineering, Centre for Life Sciences, Singapore, Singapore.
authors: Bramasta
authors: B
authors: Nugraha
authors: Xin
authors: X
authors: Hong
authors: Xuejun
authors: X
authors: Mo
authors: Looling
authors: L
authors: Tan
authors: Wenxia
authors: W
authors: Zhang
authors: Po-Mak
authors: PM
authors: Chan
authors: Chiang Huen
authors: CH
authors: Kang
authors: Yan
authors: Y
authors: Wang
authors: Lu Thong
authors: LT
authors: Beng
authors: Wanxin
authors: W
authors: Sun
authors: Deepak
authors: D
authors: Choudhury
authors: Jeffrey M
authors: JM
authors: Robens
authors: Michael
authors: M
authors: McMillian
authors: Jose
authors: J
authors: Silva
authors: Shannon
authors: S
authors: Dallas
authors: Choon-Hong
authors: CH
authors: Tan
authors: Zhilian
authors: Z
authors: Yue
authors: Hanry
authors: H
authors: Yu
mesh_terms: D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods; D002478:Cells, Cultured; D002482:Cellulose / Q000737:chemistry*; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D005690:Galactose / Q000737:chemistry*; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism; D057166:High-Throughput Screening Assays / Q000295:instrumentation / Q000379:methods*; D020100:Hydrogels / Q000737:chemistry*; D008297:Male; D008422:Materials Testing; D015394:Molecular Structure; D004364:Pharmaceutical Preparations / Q000378:metabolism; D016062:Porosity; D051381:Rats; D017208:Rats, Wistar; D018874:Spheroids, Cellular / Q000166:cytology* / Q000378:metabolism; D023822:Tissue Engineering / Q000379:methods
chemical_list: D001672:Biocompatible Materials; D020100:Hydrogels; D004364:Pharmaceutical Preparations; D002482:Cellulose; D003577:Cytochrome P-450 Enzyme System; D005690:Galactose