Assessment of Bovine Collagen Manufactured via Cellular Agriculture

Cellular agriculture offers a transformative approach to protein production by creating biologically derived alternatives to animal-sourced biomaterials. This study focuses on the comprehensive characterization of bovine collagen produced via cellular agriculture. A bovine fibroblast cell line optimized for collagen production was cultured in a custom bioreactor system to produce collagen (≥95% type I), which was extracted, purified, lyophilized, reconstituted at 8 mg/mL, and cast into self-assembling fibrillar hydrogels (Ø8 × 4 mm²) at neutral pH and 37 °C. Functional properties of this cell-based (CB) collagen were benchmarked against traditional animal-derived (AD) bovine collagen. SDS-PAGE confirmed their molecular similarity, revealing characteristic α-chains (∼120 kDa), β-dimers (∼240 kDa), and γ-trimers (∼300 kDa). Amino acid analysis showed nearly identical amino acid profiles, and differential scanning calorimetry analysis highlighted equivalent thermal stability between them. Furthermore, circular dichroism spectroscopy confirmed preservation of the triple-helical structure in the CB collagen. Mechanical testing demonstrated that CB hydrogels possessed significantly higher compressive and dynamic moduli (1260 ± 191 Pa and 6250 ± 1031 Pa, respectively) compared to the AD hydrogels (890 ± 255 Pa and 3134 ± 1857 Pa, respectively) (p < 0.05), indicating better structural integrity and shape retention. Rheological analysis supported these findings, indicated by the higher storage and loss moduli of CB hydrogels (48.40 ± 8.5 Pa and 13.20 ± 1.3 Pa, respectively) than AD hydrogels (25 ± 2.12 Pa and 10 ± 1.58 Pa, respectively) (p < 0.05). Surface wettability testing highlighted comparable hydrophilicity for CB and AD collagens. Finally, the biocompatibility of the CB hydrogels was evident from the notable increase in cultured fibroblast cell density, live cell area, and metabolic activity over 7 days. This comparative assessment underscores the functional equivalence of CB collagen with conventional animal-derived sources and offers critical insights for optimizing cellular agriculture-based production systems and hydrogel formulations for application-specific requirements in the future.