3D Cell-Based High-Content Screening (HCS) Using a Micropillar and Microwell Chip Platform.
Sang-Yun LeeIl DohDo-Hyun NamDong Woo LeePublished in: Analytical chemistry (2018)
A micropillar/microwell chip platform was applied to develop a 3D cell-based high-content screening (HCS) platform. Previously, 3D cell culture in the micropillar/microwell chip platform was only limited to cell viability measurements in a high-throughput manner. However, an HCS system could provide biological and phenotypic information which was very useful to understand complex biological functions and mechanisms of drug actions. To stain 3D cultured cells immobilized with alginate spots, we developed and optimized antibody staining procedures for 3D cultured cells. As a proof of concept, the phospho-EGFR (p-EGFR, a highly expressed receptor protein in cancer), F-actin (a protein of the actin cytoskeleton), and nuclei of 3D cultured cells were stained and analyzed after being treated with 72 different drugs. The p-EGFR inhibition of the drugs was successfully identified in the 3D cultured cells by comparing p-EGFR expression with that of F-actin and the nucleus. The p-EGFR expression levels were also measured by Western blot to verify the chip data.
Keyphrases
- high throughput
- induced apoptosis
- small cell lung cancer
- epidermal growth factor receptor
- cell cycle arrest
- tyrosine kinase
- single cell
- endoplasmic reticulum stress
- cell death
- mass spectrometry
- healthcare
- cell therapy
- signaling pathway
- machine learning
- squamous cell carcinoma
- emergency department
- small molecule
- mesenchymal stem cells
- bone marrow
- data analysis
- amino acid
- childhood cancer