Application of a 3D hydrogel-based model to replace use of animals for passaging patient-derived xenografts

Jones, Sal; Ashworth, Jennifer C.; Meakin, Marian; Collier, Pamela; Probert, Catherine; Ritchie, Alison A.; Merry, Catherine L. R.; Grabowska, Anna M.

Abstract

Purpose
This 3D in vitro cancer model for propagation of patient-derived cells, using a synthetic self-assembling peptide gel, allows the formation of a fully characterised, tailorable tumour microenvironment. Unlike many existing 3D cancer models, the peptide gel is inert, apart from molecules and motifs deliberately added or produced by cells within the model.

Methods
Breast cancer patient-derived xenografts (PDXs) were disaggregated and embedded in a peptide hydrogel. Growth was monitored by microscopic examination and at intervals, cells were extracted from the gels and passaged on into fresh gels. Passaged cells were assessed by qPCR and immunostaining techniques for the retention of characteristic markers.

Results
Breast cancer PDXs were shown to be capable of expansion over four or more passages in the peptide gel. Contaminating mouse cells were found to be rapidly removed by successive passages. The resulting human cells were shown to be compatible with a range of common assays useful for assessing survival, growth and maintenance of heterogeneity.

Conclusions
Based on these findings, the hydrogel has the potential to provide an effective and practical breast cancer model for the passage of PDXs which will have the added benefits of being relatively cheap, fully-defined and free from the use of animals or animal products. Encapsulated cells will require further validation to confirm the maintenance of cell heterogeneity, genotypes and phenotypes across passage, but with further development, including the addition of bespoke cell and matrix components of the tumour microenvironment, there is clear potential to model other cancer types.

Citation

Jones, S., Ashworth, J. C., Meakin, M., Collier, P., Probert, C., Ritchie, A. A., Merry, C. L. R., & Grabowska, A. M. (2023). Application of a 3D hydrogel-based model to replace use of animals for passaging patient-derived xenografts. In Vitro Models, 2(3-4), 99-111. https://doi.org/10.1007/s44164-023-00048-x

• Journal Article Type: Article
• Acceptance Date: Mar 15, 2023
• Online Publication Date: May 9, 2023
• Publication Date: 2023
• Deposit Date: Jun 1, 2023
• Publicly Available Date: Jun 1, 2023
• Journal: In vitro models
• Print ISSN: 2731-3433
• Electronic ISSN: 2731-3441
• Publisher: Springer Nature
• Peer Reviewed: Peer Reviewed
• Volume: 2
• Issue: 3-4
• Pages: 99-111
• DOI: https://doi.org/10.1007/s44164-023-00048-x
• Keywords: Breast cancer · In vitro model · 3D culture · Tumour microenvironment · Hydrogel · PDX
• Public URL: https://nottingham-repository.worktribe.com/output/20566967