Validation of Scinora serum-free media formulations and Xplace supplement for the development of animal-free NAMs for nanotoxicology

Fiona will build confidence in animal-free cell culture for complex in vitro models for nanotoxicity testing by evaluating and characterising non-animal derived products provided by the contract research organisation Scinora. The work will test the effects of different media on two widely used human cell lines (Calu-3 lung adenocarcinoma and THP-1 monocytes) both in monoculture and a co-culture system using a standardised operating protocol.

This award was made as part of the 2024 non-animal derived product validation grants supported with funding from the Department for Science, Innovation and Technology (DSIT).

The expanding development of new nanomaterials (NMs), particles 1000-times smaller than the width of a hair, has led to an increase in the number of materials which require robust toxicity testing to ensure safety of exposed populations. Traditionally testing has relied on exposing rodents to materials as a surrogate for humans however in recent years there has been a shift towards the development of alternative Novel Approach Methodologies (NAMs) including in silico, in chemico and in vitro methods, to reduce and replace the use of animals in toxicology research. The shift to wider adoption of NAMs will have a seismic impact on the reduction of animal numbers used directly in toxicology studies, however the use of animals to generate reagents and supplements currently required in the standard operating protocols (SOPs) for many NAMs remains as a significant burden. The replacement of animal-derived products incorporated into the NAMs protocols has received less attention to date but as NAMs progress towards validation it is timely to consider how to optimise the NAM under development without the need for animal-derived products.

The overall aim of this project is to assess the impact of replacing animal-derived reagents with human or synthetic reagents when designing SOPs for the culture and use of cell models for NM toxicity testing, in response to the current drive towards adopting animal-free approaches in toxicology. Specifically, chemically defined media and human-derived supplements developed by Scinora GmBH will be validated for the culture of airway epithelial cells and immune cells, namely Calu-3 bronchial epithelial cells and THP-1 monocytes, advancing the development of airway NAMs for the hazard assessment of NMs.

A systematic strategy has been designed to select the most appropriate media formulation to support cell line proliferation followed by comprehensive characterisation evaluating the impact of the alternative media formulation on cell phenotype and functionality, including validation of the use of Scinora media supported cell culture for NM toxicity testing. Through transparent reporting of results and the dissemination of an alternative SOP for Calu-3 and THP-1 culture and exposure to NMs the outputs of this project will promote the removal of animal-derived products from nanotoxicology NAMs thereby reducing ethical concerns and increasing the human relevance of toxicity testing of novel and advanced inhalable NM currently on the market and in future development.