3D Bioprinting of High-Grade Serous Ovarian Cancer Cells: Workflow for Gelatin-Based Bioink Formulation with...

Ovarian cancer (OC) is one of the leading causes of cancer-related mortality in women, primarily due to the late-stage diagnosis and limited therapeutic efficacy. Three-dimensional (3D) bioprinting enables the fabrication of cell-laden scaffolds that better recapitulate the native spatial organization and may support the investigation of tumor cell behavior. However, high-grade serous ovarian cancer (HG-SOC) cell subtypes remain largely unexplored in bioprinting, and it remains unclear whether a different scaffold composition is required to provide an optimal niche for each subtype. To address these gaps, we developed a 3D bioprinting workflow based on methacrylated gelatin (GelMA) and a hybrid methacrylated hyaluronic acid/methacrylated gelatin formulation (HAGel) to generate constructs containing KURAMOCHI and OVCAR3 HG-SOC cells, alongside SKOV3 as a reference epithelial OC model. Bioink formulations were optimized to achieve reproducible extrusion printing and stable construct formation. Rheological characterization showed a higher storage modulus for HAGel scaffold (approximately 2-fold higher than GelMA), while both formulations remained within a stiffness range suitable for cell culture. GelMA exhibited higher swelling and a broader printability window, whereas HAGel showed reduced water uptake and required more restricted processing conditions to ensure stable bioprinting. Cellular responses were evaluated in terms of viability, proliferation, morphology, and expression of hyaluronic-acid-associated markers (e.g., CD44, MMP-2, and MMP-14). Postprinting viability remained high throughout culture (approximately 80-99% after 14 days), with outcomes depending on both cell line and hydrogel formulation. KURAMOCHI and SKOV3 cells exhibited higher viability and cell density in HAGel, whereas OVCAR3 cells performed better in GelMA, consistent with CD44 expression and MMP trends, reflecting subtype-dependent interactions with the scaffold composition. These findings demonstrate the feasibility of bioprinting HG-SOC cells and show how bioink formulations may contribute to cell-line-dependent response, paving the way for a rational design of constructs for advanced OC in vitro platforms.