Modular design and development of PEG-based hydrogel 3D systems for human intestinal organoid culture

Sandy, Joseph Daniel (2023) Modular design and development of PEG-based hydrogel 3D systems for human intestinal organoid culture. Doctoral thesis, University of East Anglia.

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Abstract

Human intestinal organoids (HIO) are a powerful 3D, platform for use in research and clinical environments. Due to their organotypic morphology and resemblance to the native colonic epithelium in structure and function, HIO’s have been utilized to model complex disease pathologies. The full potential of HIO remains unexplored, due to the reliance of 3D organoid growth within an ill-defined, tumor-derived basement membrane extract known commercially as Matrigel. Matrigel is derived from a rat-sarcoma cell line, which presents the risk of immunogen and pathogen transfer. This has provided the need for a chemically defined synthetic hydrogel which supports the 3D culture of HIO. Research has indicated that defined hydrogels can be optimised with various extracellular matrix (ECM) components and adhesive ligands which facilitate organoid culture to a similar standard as Matrigel. Additionally, the role of matrix environments such as stiffness have an impact on mechano-transduction pathways influencing stem cell biology and potentially organoid development. Growth of human colonic organoids has not been fully achieved in a defined environment and there remains the potential for a fully defined and optimised hydrogel designed specifically for optimal HIO culture. We demonstrate HIO growth can be achieved in a synthetic, fully defined hydrogel, based on the reaction between four-armed, thiol and maleimide-terminated poly(ethylene glycol) (PEG) macromers. We identify that PEG hydrogels are a suitable candidate for continued optimisation towards HIO culture due to their ability to support organoid growth and viability to a limited capacity in a non-functionalised state. the sub-optimal organoid morphology and overall change in organoid cross-sectional area over 7 days in non-functional PEG gels can be improved by the inclusion of the biologically relevant ECM proteins such as laminin and replicated with fully defined synthetic peptides. This work provides insight into the mechanical and biochemical requirements for colonic human intestinal organoid culture.

Item Type:	Thesis (Doctoral)
Faculty \ School:	Faculty of Science > School of Pharmacy (former - to 2024)
Depositing User:	Nicola Veasy
Date Deposited:	12 Nov 2024 15:14
Last Modified:	12 Nov 2024 15:14
URI:	https://ueaeprints.uea.ac.uk/id/eprint/97658
DOI:

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