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Open AccessTechnical Note

A High Throughput Apoptosis Assay using 3D Cultured Cells

by Sang-Yun Lee 1,2, Il Doh 3,* and Dong Woo Lee 4,*
Department of Health sciences and Technology, SAIHST, Sungkyunkwan University, Seoul 06351, Korea
Medical & Bio Device (MBD), Suwon 16229, Korea
Center for Medical Convergence Metrology, Korea Research Institute of Standards and Science, Daejeon 34113, Korea
Department of Biomedical Engineering, Konyang University, Daejon 35365, Korea
Authors to whom correspondence should be addressed.
Academic Editors: Anton Simeonov and Zhihao Zhuang
Molecules 2019, 24(18), 3362;
Received: 15 July 2019 / Revised: 6 September 2019 / Accepted: 10 September 2019 / Published: 16 September 2019
(This article belongs to the Special Issue High Throughput Screening II)
A high throughput apoptosis assay using 3D cultured cells was developed with a micropillar/microwell chip platform. Live cell apoptosis assays based on fluorescence detection have been useful in high content screening. To check the autofluorescence of drugs, controls (no caspase-3/7 reagent in the assay) for the drugs are necessary which require twice the test space. Thus, a high throughput capability and highly miniaturized format for reducing reagent usage are necessary in live cell apoptosis assays. Especially, the expensive caspase-3/7 reagent should be reduced in a high throughput screening system. To solve this issue, we developed a miniaturized apoptosis assay using micropillar/microwell chips for which we tested seventy drugs (six replicates) per chip and reduced the assay volume to 1 µL. This reduced assay volume can decrease the assay costs compared to the 10–40 µL assay volumes used in 384 well plates. In our experiments, among the seventy drugs, four drugs (Cediranib, Cabozatinib, Panobinostat, and Carfilzomib) induced cell death by apoptosis. Those results were confirmed with western blot assays and proved that the chip platform could be used to identify high potency apoptosis-inducing drugs in 3D cultured cells with alginate. View Full-Text
Keywords: apoptosis assay; caspase-3/7; 3D cell culture; cell encapsulation; high-throughput screening apoptosis assay; caspase-3/7; 3D cell culture; cell encapsulation; high-throughput screening
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Lee, S.-Y.; Doh, I.; Lee, D.W. A High Throughput Apoptosis Assay using 3D Cultured Cells. Molecules 2019, 24, 3362.

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