Validating methods to cryopreserve, store and revive human precision cut tissue slices for disease modelling and drug testing

Importance: Organ fibrosis is the pathological consequence of tissue damage and inflammation and is estimated to underlie ~45% of deaths in the western world [1]. Treatment options for fibrosis are limited, highlighting an urgent need to better understand the disease biology to identify new anti-fibrotic medicines and test novel therapies.

Simple cell culture systems and animal models fail to faithfully recreate human disease, therefore have limited utility to study inflammation and fibrosis biology and test novel therapies. Human precision cut tissue slices (hPCTS) retain the architecture, cellular heterogeneity, cell-cell and cell-matrix communications of an organ and represent a robust non-animal technology to model human disease and perform drug efficacy or toxicity testing [2].

However, access to human tissue for research, and the necessary infrastructure and technical expertise of hPCTS methodology is currently limited to specialist research teams. This presents a major barrier for the scientific community to adopt hPCTS, as non-animal replacement technology for translational research and drug discovery programs.  In addition, hPCTS experiments require fresh tissue, which can present logistical and capacity issues.

A solution to overcome these barriers and maximise human tissue use, is to cryopreserve hPCTS and generate banks of highly characterised frozen hPCTS for “on demand” use in disease-modelling experiments and drug screens.

Vision: Our team have pioneered the use of lung, kidney and liver hPCTS to model tissue inflammation & fibrosis and assess drug efficacy [3-8], and our vision is to increase the 3Rs impact of hPCTS models in our group and promote their adoption by the wider research community and Pharma.

To achieve this vision the focus of this studentship proposal is to establish robust methods to cryopreserve and revive hPCTS and then validate their use to model disease and evaluate drug efficacy.

Scientific objectives:

1. Develop methods to cryopreserve and revive human precision cut lung (hPCLuS) and kidney (hPCKS) slices for ex-vivo culture.
2. Evaluation of cryopreserved hPCLuS and hPCKS for modelling of inflammation and fibrosis and for drug efficacy testing.
3. Expansion of cryopreservation methods to human liver (hPCLS) and heart (hPCHS) slices

Why we will succeed: Our innovative team are passionate about delivering this research and have the technical expertise, equipment, tissue access and research environment needed to successfully deliver this clinically relevant, 3Rs impactful project. By publishing detailed methods and establishing a hPCTS bank at Newcastle for the community, we can help support the replacement of in vivo models with hPCTS.

[1] Wynn, T.A., Fibrotic disease and the T(H)1/T(H)2 paradigm. Nat Rev Immunol, 2004. 4(8): p. 583-94.
[2] Oakley, F., et al., Implementation of pre-clinical methodologies to study fibrosis and test anti-fibrotic therapy. Curr Opin Pharmacol, 2019. 49: p. 95-101.