Semi-Synthetic Click-Gelatin Hydrogels as Tunable Platforms for 3D Cancer Cell Culture

• Published in: Gels Vol. 8; no. 12; p. 821
• Main Authors: Hipwood, Luke, Clegg, Julien, Weekes, Angus, Davern, Jordan W, Dargaville, Tim R, Meinert, Christoph, Bock, Nathalie
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 12.12.2022; MDPI
• Subjects: 3D cancer model; Amino acids; Biological products; Biomedical materials; Cancer; Cell culture; click chemistry; Collagen; Crosslinked polymers; Crosslinking; Differentiation (biology); Ethylene glycol; extracellular matrix; Gelatin; Hydrogels; Mechanical properties; Polyethylene glycol; Raw materials; Rheology; Viscosity; Australia; New York; Massachusetts; United States > US; click chemistry; hydrogels; 3D cancer model; cell culture; gelatin; extracellular matrix
• ISSN: 2310-2861; 2310-2861
• DOI: 10.3390/gels8120821

Basement membrane extracts (BME) derived from Engelbreth-Holm-Swarm (EHS) mouse sarcomas such as Matrigel remain the gold standard extracellular matrix (ECM) for three-dimensional (3D) cell culture in cancer research. Yet, BMEs suffer from substantial batch-to-batch variation, ill-defined composition, and lack the ability for physichochemical manipulation. Here, we developed a novel 3D cell culture system based on thiolated gelatin (Gel-SH), an inexpensive and highly controlled raw material capable of forming hydrogels with a high level of biophysical control and cell-instructive bioactivity. We demonstrate the successful thiolation of gelatin raw materials to enable rapid covalent crosslinking upon mixing with a synthetic poly(ethylene glycol) (PEG)-based crosslinker. The mechanical properties of the resulting gelatin-based hydrogels were readily tuned by varying precursor material concentrations, with Young's moduli ranging from ~2.5 to 5.8 kPa. All hydrogels of varying stiffnesses supported the viability and proliferation of MDA-MB-231 and MCF-7 breast cancer cell lines for 14 and 21 days of cell culture, respectively. Additionally, the gelatin-based hydrogels supported the growth, viability, and osteogenic differentiation of patient-derived preosteoblasts over 28 days of culture. Collectively, our data demonstrate that gelatin-based biomaterials provide an inexpensive and tunable 3D cell culture platform that may overcome the limitations of traditional BMEs.