Valorizing watermelon rind waste into Plant-Based scaffolds for Proof-of-Concept cultivated meat production in a dynamic Fixed-Bed bioreactor

Cultivated meat (CM) biomanufacturing faces major challenges in cost and in scaling adherent-cell processes from the laboratory toward pilot and industrial production. This study presents a proof-of-concept (POC) in which watermelon rind, an agricultural waste by-product, was valorized into decellularized plant scaffolds (vDPS) and evaluated in a dynamic fixed-bed reactor (FBR). By replacing Triton® X-100 (1% v/v) with Tween-20 (10% v/v), we achieved effective decellularization, reducing residual DNA to below 25 ng/mg while eliminating the use of the toxic Triton X-100, making the scaffolds more suitable for CM applications. The resulting scaffolds exhibited high porosity (85.6%) and a Young's modulus of 36.8 ± 5.9 kPa, confirming their suitability for use as CM scaffolds. For dynamic bioreactor evaluation, the scaffolds were coated with Corning® Synthemax® II SC substrate and cultured with murine myoblast C2C12 cells under the process control capabilities of the Corning Ascent® fixed bed reactor (FBR) system. Over seven days, cells attached, proliferated, and differentiated; relative to static culture, the dynamic condition showed higher metabolic activity and denser cell coverage. Myogenic differentiation was confirmed by Myosin heavy chain and Desmin expression, and cross-sectional imaging showed cell penetration into the scaffold. The prototypes remained structurally stable after cooking. Overall, this work demonstrates the feasibility of integrating waste-valorized scaffolds into a dynamic FBR system, providing a POC foundation for future scale-up studies.