Preparation of a User-Defined Peptide Gel for Controlled 3D Culture Models of Cancer and Disease

Abstract

There is a growing awareness that cells grown in 3D better model in vivo behavior than those grown in 2D. In this protocol, we describe a simple and tunable 3D hydrogel, suitable for culturing cells and tissue in a setting that matches their native environment. This is particularly important for researchers investigating the initiation, growth, and treatment of cancer where the interaction between cells and their local extracellular matrix is a fundamental part of the model. Moving to 3D culture can be challenging and is often associated with a lack of reproducibility due to high batch-to-batch variation in animal-derived 3D culture matrices. Similarly, handling issues can limit the usefulness of synthetic hydrogels. In response to this need, we have optimized a simple selfassembling peptide gel, to enable the culture of relevant cell line models of cancer and disease, as well as patient-derived tissue/cells. The gel itself is free from matrix components, apart from those added during encapsulation or deposited into the gel by the encapsulated cells. The mechanical properties of the hydrogels can also be altered independent of matrix addition. It, therefore, acts as a 'blank slate' allowing researchers to build a 3D culture environment that reflects the target tissue of interest and to dissect the influences of mechanical forces and/or biochemical control of cell behavior independently.

OptoRheo: Simultaneousin situmicro-mechanical sensing and imaging of live 3D biological systems

“…The precursor and gel preparation method was followed as previously published 57 . A commercially available peptide preparation in powder form was used as the source of the octapeptide gelator (Pepceuticals UK, FEFEFKFK, Phe-Glu-Phe-Glu-Phe-Lys-Phe-Lys).…”

Recent Advances in Multicellular Tumor Spheroid Generation for Drug Screening

“…Biomaterials for gel-type 3D cell culture are derived from natural and artificially modified materials [69]. Since gels allow independent control of matrix and ECM functionalization, Ashworth et al, (2020) demonstrated a self-assembling peptide gel for designing the 3D cell culture [70]. By controlling stiffness through peptide concentration and pH condition, the peptide gel was fully utilized with further experiments on culturing breast cancer.…”