A novel 3D polycaprolactone high-throughput system for evaluation of toxicity in normoxia and hypoxia. (2021)

• Record Type: Journal Article
• Authors: Malakpour-Permlid, Atena; Oredsson, Stina
• Is Part Of: Toxicology reports. Volume 8(2021)
• Page Start: 627
• Page End: 635
• Publication Date: 2021
• Subjects: 2D two-dimensional -- 3D three-dimensional -- CSCs cancer stem cells -- DHHS donor herd horse serum -- ECM extracellular matrix -- FBS fetal bovine serum -- HDFs human dermal fibroblasts -- HTS high-throughput -- PCL polycaprolactone -- SAEC salinomycin analogue 20-ethyl carbonate-Na; 3D high-throughput screening -- Polycaprolactone fibre network -- Breast cancer cells -- Paclitaxel -- Multi-well plates -- Hypoxia; Toxicology -- Periodicals; Clinical toxicology -- Periodicals; Drug-Related Side Effects and Adverse Reactions; Hazardous Substances; Poisoning; Toxicology; Electronic journals; Periodicals; Periodicals; 571.9505
• DOI: 10.1016/j.toxrep.2021.03.015
• Languages: English
• ISSNs: 2214-7500

Abstract

Highlights: Development of 3D polycaprolactone-based high throughput 96-well plate assay for pre-clinical compound screening. Normal fibroblasts and JIMT-1 breast cancer cells infiltrate throughout the depth of the 200 μm thick 3D fibre network. The toxicity is compared between cells grown in flat 2D and in 3D 96-well plates incubated in normoxia or hypoxia. Dose-response curves show that the two cell lines are more sensitive to treatment when grown in 2D compared to 3D. Abstract: Two-dimensional (2D) culturing of cancer cells has been indispensable for the development of anti-cancer drugs. Drug development, however, is lengthy and costly with a high attrition rate, calling to mind that 2D culturing does not mimic the three-dimensional (3D) tumour microenvironment in vivo . Thus, began the development of 3D culture models for cancer research. We have constructed a 3D 96-well plate using electrospun fibres made of biocompatible polycaprolactone (PCL). Finely-cut PCL fibre pieces in water/ethanol solution was pipetted to the wells of hydrophobic 96-well plates. A fibrous network of approximately 200 μm thickness and high porosity was formed after crosslinking and drying. Human JIMT-1 breast cancer cells or fibroblasts were seeded into the network. Confocal microscopy shows that the cells grow throughout the fibre network. The toxicity of paclitaxel and an experimental salinomycin analogue was assessed and compared in 2D and 3D cultures incubated under conditions of normoxia and hypoxia often found in tumours. The toxicity of both compounds is lower when the cells are cultured in 3D compared to 2D in either normoxia or hypoxia. We conclude that our 96-well assay is a cost-efficient tool that may be used for high-throughput pre-clinical screening of potential anti-cancer compounds.