2024 AIChE Annual Meeting
(413g) A Novel Hydrogel Matrix for Three-Dimensional Culturing of Breast Cancer Cells
Authors
Ashraf Al-Goraee - Presenter, University of North Dakota
Ali Alshami, University of North Dakota
The development of cancer treatment can be accelerated by coupling regenerative and personalized medicine. In-vitro modeling of cancerous tumors combines both aspects, holding the promise of more effective, patient-tailored therapeutic strategies and more efficient laboratory screening of drugs. Additionally, this approach provides a safe, cost-effective platform to develop new formulas for anticancer drugs and determine the exact bioactivity against cancer cells. Three-dimensional cell culture has outperformed the traditional monolayer approach in mimicking the structural and functional properties of the native extracellular matrix (ECM). Different cell-cell and cell-ECM activities are promoted when cells are seeded in 3D replicating morphological and physiological cues of the native ECM. Tunable materials such as hydrogels provide a reliable niche for 3D cell culture due to their adaptable mechanical properties, high water content, controlled porosity, permeability to oxygen and nutrients, and intrinsic biocompatibility. All these properties are pivotal for crucial applications such as in-vitro cancer modeling and laboratory screening of drugs. In this work, a new hydrogel matrix was developed to synthesize an interpenetrating network hydrogel of synthetic and natural polymers. The reversible physical crosslinking of freeze-thaw cycles was utilized to establish a hydrogel matrix with different material porosity and stiffness. Thorough material characterization was applied to evaluate the homogeneity, stability, functionality, and tunability of the hydrogel. The utilized techniques included FTIR, TGA, DSC, mechanical testing, and SEM. The developed hydrogels showed reliable and tailored mechanical properties, thermal stability up to 200 oC, and freeze-thaw cycles related pore size. After the comprehensive material characterization, the hydrogels were subjected to essential biocompatibility testing such as media incubation and pH monitoring. After that, the cell lines MCF7 and MCF10A were separately seeded within three different stiffness levels labeled as soft, medium, and hard. The cells were rapidly growing and successfully formed good quality and well-defined spheroids. Phenotype and immunofluorescence assays were performed to characterize the spheroids at the three stiffnesses and assess their viability, morphologies, cell-cell and cell-ECM interactions. The DAPI staining and E-cadherin expression confirmed the cell's viability and cell-cell adhesion within the spheroids respectively. Finally, intact and high-yield RNA was successfully extracted for subsequent analyses ensuring the hydrogel's ability to preserve the nuclear content after cell lysis.