Simple in-house fabrication of microwells for generating uniform hepatic multicellular cancer aggregates and discovering novel therapeutics

Chiao Yi Chiu, Ying Chi Chen, Kuang Wei Wu, Wen Chien Hsu, Hong Ping Lin, Hsien Chang Chang, Yung Chun Lee, Yang Kao Wang, Ting Yuan Tu

Research output: Contribution to journalArticle

Abstract

Three-dimensional (3D) cell culture models have become powerful tools because they better simulate the in vivo pathophysiological microenvironment than traditional two-dimensional (2D) monolayer cultures. Tumor cells cultured in a 3D system as multicellular cancer aggregates (MCAs) recapitulate several critical in vivo characteristics that enable the study of biological functions and drug discovery. The microwell, in particular, has emerged as a revolutionary technology in the generation of MCAs as it provides geometrically defined microstructures for culturing size-controlled MCAs amenable for various downstream functional assays. This paper presents a simple and economical microwell fabrication methodology that can be conveniently incorporated into a conventional laboratory setting and used for the discovery of therapeutic interventions for liver cancer. The microwells were 400-700 μm in diameter, and hepatic MCAs (Huh-7 cells) were cultured in them for up to 5 days, over which time they grew to 250-520 μm with good viability and shape. The integrability of the microwell fabrication with a high-throughput workflow was demonstrated using a standard 96-well plate for proof-of-concept drug screening. The IC50 of doxorubicin was determined to be 9.3 μM under 2D conditions and 42.8 μM under 3D conditions. The application of photothermal treatment was demonstrated by optimizing concanavalin A-FITC conjugated silica-carbon hollow spheres (SCHSs) at a concentration of 500:200 μg/mL after a 2 h incubation to best bind with MCAs. Based on this concentration, which was appropriate for further photothermal treatment, the relative cell viability was assessed through exposure to a 3 W/cm2 near-infrared laser for 20 min. The relative fluorescence intensity showed an eight-fold reduction in cell viability, confirming the feasibility of using photothermal treatment as a potential therapeutic intervention. The proposed microwell integration is envisioned to serve as a simple in-house technique for the generation of MCAs useful for discovering therapeutic modalities for liver cancer treatment.

Original languageEnglish
Article number3308
JournalMaterials
Volume12
Issue number20
DOIs
Publication statusPublished - 2019 Oct 1

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Fabrication
Cells
Cell culture
Liver
Oncology
Infrared lasers
Fluorescein-5-isothiocyanate
Concanavalin A
Silicon Dioxide
Doxorubicin
Tumors
Monolayers
Assays
Screening
Carbon
Fluorescence
Silica
Throughput
Microstructure
Pharmaceutical Preparations

All Science Journal Classification (ASJC) codes

  • Materials Science(all)

Cite this

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title = "Simple in-house fabrication of microwells for generating uniform hepatic multicellular cancer aggregates and discovering novel therapeutics",
abstract = "Three-dimensional (3D) cell culture models have become powerful tools because they better simulate the in vivo pathophysiological microenvironment than traditional two-dimensional (2D) monolayer cultures. Tumor cells cultured in a 3D system as multicellular cancer aggregates (MCAs) recapitulate several critical in vivo characteristics that enable the study of biological functions and drug discovery. The microwell, in particular, has emerged as a revolutionary technology in the generation of MCAs as it provides geometrically defined microstructures for culturing size-controlled MCAs amenable for various downstream functional assays. This paper presents a simple and economical microwell fabrication methodology that can be conveniently incorporated into a conventional laboratory setting and used for the discovery of therapeutic interventions for liver cancer. The microwells were 400-700 μm in diameter, and hepatic MCAs (Huh-7 cells) were cultured in them for up to 5 days, over which time they grew to 250-520 μm with good viability and shape. The integrability of the microwell fabrication with a high-throughput workflow was demonstrated using a standard 96-well plate for proof-of-concept drug screening. The IC50 of doxorubicin was determined to be 9.3 μM under 2D conditions and 42.8 μM under 3D conditions. The application of photothermal treatment was demonstrated by optimizing concanavalin A-FITC conjugated silica-carbon hollow spheres (SCHSs) at a concentration of 500:200 μg/mL after a 2 h incubation to best bind with MCAs. Based on this concentration, which was appropriate for further photothermal treatment, the relative cell viability was assessed through exposure to a 3 W/cm2 near-infrared laser for 20 min. The relative fluorescence intensity showed an eight-fold reduction in cell viability, confirming the feasibility of using photothermal treatment as a potential therapeutic intervention. The proposed microwell integration is envisioned to serve as a simple in-house technique for the generation of MCAs useful for discovering therapeutic modalities for liver cancer treatment.",
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Simple in-house fabrication of microwells for generating uniform hepatic multicellular cancer aggregates and discovering novel therapeutics. / Chiu, Chiao Yi; Chen, Ying Chi; Wu, Kuang Wei; Hsu, Wen Chien; Lin, Hong Ping; Chang, Hsien Chang; Lee, Yung Chun; Wang, Yang Kao; Tu, Ting Yuan.

In: Materials, Vol. 12, No. 20, 3308, 01.10.2019.

Research output: Contribution to journalArticle

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AU - Chiu, Chiao Yi

AU - Chen, Ying Chi

AU - Wu, Kuang Wei

AU - Hsu, Wen Chien

AU - Lin, Hong Ping

AU - Chang, Hsien Chang

AU - Lee, Yung Chun

AU - Wang, Yang Kao

AU - Tu, Ting Yuan

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