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Title: Three-dimensional aggregated spheroid model of hepatocellular carcinoma using a 96-pillar/well plate
Authors: Lee, Sang-Yun
Teng, Yvonne
Son, Miseol
Ku, Bosung
Hwang, Hyun Ju
Tergaonkar, Vinay 
Chow, Pierce Kah-Hoe 
Lee, Dong Woo
Nam, Do-Hyun
Keywords: 3D cell culture
96-pillar/well plate
Cancer spheroids in Matrigel
Hepatocellular carcinoma cell line
High-throughput screening
In vitro extracellular matrix remodel-ing
Issue Date: 16-Aug-2021
Publisher: MDPI AG
Citation: Lee, Sang-Yun, Teng, Yvonne, Son, Miseol, Ku, Bosung, Hwang, Hyun Ju, Tergaonkar, Vinay, Chow, Pierce Kah-Hoe, Lee, Dong Woo, Nam, Do-Hyun (2021-08-16). Three-dimensional aggregated spheroid model of hepatocellular carcinoma using a 96-pillar/well plate. Molecules 26 (16) : 4949. ScholarBank@NUS Repository.
Rights: Attribution 4.0 International
Abstract: A common method of three-dimensional (3D) cell cultures is embedding single cells in Matrigel. Separated cells in Matrigel migrate or grow to form spheroids but lack cell-to-cell interaction, which causes difficulty or delay in forming mature spheroids. To address this issue, we proposed a 3D aggregated spheroid model (ASM) to create large single spheroids by aggregating cells in Matrigel attached to the surface of 96-pillar plates. Before gelling the Matrigel, we placed the pillar inserts into blank wells where gravity allowed the cells to gather at the curved end. In a drug screening assay, the ASM with Hepatocellular carcinoma (HCC) cell lines showed higher drug resistance compared to both a conventional spheroid model (CSM) and a two-dimensional (2D) cell culture model. With protein expression, cytokine activation, and penetration analysis, the ASM showed higher expression of cancer markers associated with proliferation (p-AKT, p-Erk), tight junction formation (Fibronectin, ZO-1, Occludin), and epithelial cell identity (E-cadherin) in HCC cells. Furthermore, cytokine factors were increased, which were associated with immune cell recruitment/activation (MIF-3?), extracellular matrix regulation (TIMP-2), cancer interaction (IL-8, TGF-?2), and angiogenesis regulation (VEGF-A). Compared to CSM, the ASM also showed limited drug penetration in doxorubicin, which appears in tissues in vivo. Thus, the proposed ASM better recapitulated the tumor microenvironment and can provide for more instructive data during in vitro drug screening assays of tumor cells and improved prediction of efficacious drugs in HCC patients. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.
Source Title: Molecules
ISSN: 1420-3049
DOI: 10.3390/molecules26164949
Rights: Attribution 4.0 International
Appears in Collections:Staff Publications

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