title: In vitro co-culture model of medulloblastoma and human neural stem cells for drug delivery assessment.
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.
authors.affiliation: School of Pharmacy, University of Nottingham, Nottingham, UK. Electronic address: paxdi@nottingham.ac.uk.
authors.forename: Delyan P
authors.initials: DP
authors.lastname: Ivanov
authors.affiliation: Medical School, Queens Medical Centre, University of Nottingham, Nottingham, UK. Electronic address: terry.parker@nottingham.ac.uk.
authors.forename: Terry L
authors.initials: TL
authors.lastname: Parker
authors.affiliation: Children's Brain Tumour Research Centre, Queens Medical Centre, University of Nottingham, Nottingham, UK. Electronic address: david.walker@nottingham.ac.uk.
authors.forename: David A
authors.initials: DA
authors.lastname: Walker
authors.affiliation: School of Pharmacy, University of Nottingham, Nottingham, UK. Electronic address: cameron.alexander@nottingham.ac.uk.
authors.forename: Cameron
authors.initials: C
authors.lastname: Alexander
authors.affiliation: AstraZeneca, Silk Road, Macclesfield, UK. Electronic address: Marianne.Ashford@astrazeneca.com.
authors.forename: Marianne B
authors.initials: MB
authors.lastname: Ashford
authors.affiliation: AstraZeneca, Silk Road, Macclesfield, UK. Electronic address: Paul.Gellert@astrazeneca.com.
authors.forename: Paul R
authors.initials: PR
authors.lastname: Gellert
authors.affiliation: School of Pharmacy, University of Nottingham, Nottingham, UK. Electronic address: m.garnett@nottingham.ac.uk.
authors.forename: Martin C
authors.initials: MC
authors.lastname: Garnett
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D001932:Brain Neoplasms / Q000473:pathology; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D018920:Coculture Techniques / Q000379:methods*; D004354:Drug Screening Assays, Antitumor; D005047:Etoposide / Q000494:pharmacology; D006801:Humans; D008527:Medulloblastoma / Q000473:pathology; D008954:Models, Biological; D058953:Neural Stem Cells / Q000166:cytology* / Q000187:drug effects; D018874:Spheroids, Cellular / Q000187:drug effects*
chemical_list: D000970:Antineoplastic Agents; D005047:Etoposide
keywords: Cancer drug screening in 3D; Co-culture spheroid; Human stem cell neurosphere; Medulloblastoma; Three-dimensional tumor model