In vitro co-culture model of medulloblastoma and human neural stem cells for drug delivery assessment

Ivanova, Delyan P.; Parker, Terry L.; Walker, David A.; Alexander, Cameron; Ashford, Marianne B.; Gellert, Paul R.; Garnett, Martin C.

Abstract

Physiologically relevant in vitro models can serve as biological analytical platforms for testing novel treatments and drug delivery systems. We describe the first steps in the development of a 3D human brain tumour co-culture model that includes the interplay between normal and tumour tissue along with nutrient gradients, cell-cell and cell-matrix interactions. The human medulloblastoma cell line UW228-3 and human foetal brain tissue were marked with two supravital fluorescent dyes (CDCFDASE, Celltrace Violet) and cultured together in ultra-low attachment 96-well plates to form reproducible single co-culture spheroids (d = 600 μm, CV% = 10%). Spheroids were treated with model cytotoxic drug etoposide (0.3–100 μM) and the viability of normal and tumour tissue quantified separately using flow cytometry and multiphoton microscopy. Etoposide levels of 10 μM were found to maximise toxicity to tumours (6.5% viability) while stem cells maintained a surviving fraction of 40%. The flexible cell marking procedure and high-throughput compatible protocol make this platform highly transferable to other cell types, primary tissues and personalised screening programs. The model's key anticipated use is for screening and assessment of drug delivery strategies to target brain tumours, and is ready for further developments, e.g. differentiation of stem cells to a range of cell types and more extensive biological validation.

Citation

Ivanova, D. P., Parker, T. L., Walker, D. A., Alexander, C., Ashford, M. B., Gellert, P. R., & Garnett, M. C. (2015). In vitro co-culture model of medulloblastoma and human neural stem cells for drug delivery assessment. Journal of Biotechnology, 205, 3-13. https://doi.org/10.1016/j.jbiotec.2015.01.002

• Journal Article Type: Article
• Acceptance Date: Jan 5, 2015
• Online Publication Date: Jan 12, 2015
• Publication Date: Jul 10, 2015
• Deposit Date: Sep 20, 2017
• Publicly Available Date: Jan 21, 2019
• Journal: Journal of Biotechnology
• Electronic ISSN: 1873-4863
• Publisher: Elsevier
• Peer Reviewed: Peer Reviewed
• Volume: 205
• Pages: 3-13
• DOI: https://doi.org/10.1016/j.jbiotec.2015.01.002
• Public URL: https://nottingham-repository.worktribe.com/output/1110552
• Publisher URL: https://www.sciencedirect.com/science/article/pii/S0168165615000115
• PMID: 00035455
• Additional Information: This article is maintained by: Elsevier; Article Title: In vitro co-culture model of medulloblastoma and human neural stem cells for drug delivery assessment; Journal Title: Journal of Biotechnology; CrossRef DOI link to publisher maintained version: https://doi.org/10.1016/j.jbiotec.2015.01.002; Content Type: article; Copyright: Copyright © 2015 The Authors. Published by Elsevier B.V.
• Contract Date: Jan 21, 2019