392: Key Role of Scaffold Design in Cultivated Meat Production: Development of Chitosan and Gelatin-Based Scaffolds for Improved Cell Growth and Tissue Formation

Introduction

Cultivated meat, produced by culturing animal muscle and fat cells, offers a sustainable alternative to traditional meat. Its success depends on forming a three-dimensional (3D) tissue structure that mimics the texture and quality of conventional meat. Scaffolds play a crucial role in supporting cell growth, differentiation, and tissue formation. This study focuses on developing chitosan (CS) and gelatin-based scaffolds for cultivated meat production. CS, derived from chitin, is biocompatible and cost-effective but lacks sufficient mechanical strength. In contrast, collagen mimics the extracellular matrix (ECM) and provides superior mechanical properties. However, its high cost makes gelatin a viable alternative. This study aims to develop porous fiber-based scaffolds by combining gelatin and CS using tensile spinning technology, replicating ECM’s structural and mechanical properties while promoting cell adhesion.

Methods

The rheological properties of the fiber solutions were analyzed to investigate their flow behavior and viscosity. The morphology of the scaffolds was observed using scanning electron microscopy (SEM), and their chemical composition was analyzed by Fourier-transform infrared (FT-IR) spectroscopy, X-ray diffraction (XRD), and differential scanning calorimetry (DSC). In vitro characterization included evaluations of swelling, degradation, and mechanical properties. The cellular response was evaluated by culturing L6 rat myoblasts on the scaffolds and assessing cell adhesion and viability using confocal microscopy.

Results

The addition of CS to the gelatin solution resulted in an increase in viscosity, which became more pronounced as the CS concentration increased due to enhanced electrostatic interactions. SEM images showed that higher CS concentrations improved fiber alignment and reduced fiber diameter, while the scaffold density decreased due to thinner fibers and larger inter-fiber spaces. FTIR analysis confirmed crosslinking between gelatin and CS, while XRD showed reduced crystallinity, facilitating better fiber formation. DSC analysis revealed that the melting point and enthalpy increased with CS concentration but decreased at higher concentrations, indicating that excessive crosslinking weakened thermal properties. Cellular analysis showed that, consistent with other results, mechanical properties and cell adhesion improved with increasing CS concentration, but excessive CS negatively impacted both mechanical properties and cell adhesion.

Significance

Optimizing CS concentration is crucial for enhancing the scaffold’s structural and mechanical properties while maintaining favorable cell culture conditions.

Authors: Yourim Oh, and Jin-Kyu Rhee