Upscaled Skeletal Muscle Engineered Tissue with In Vivo Vascularization and Innervation Potential

Engineering functional tissues of clinically relevant size (in mm-scale) in vitro is still a challenge tissue engineering due to low oxygen diffusion and lack vascularization. To address these limitations, perfusion bioreactor was used generate contractile engineered muscles 3 mm-thickness 8 mm-diameter. This study aimed upscale the process 50 mm diameter by combining murine skeletal myoblasts (SkMbs) with human adipose-derived stromal vascular fraction (SVF) cells, providing high neuro-vascular potential vivo. SkMbs were cultured on type-I-collagen scaffold (co-culture) or without (monoculture) SVF. Large-scale muscle-like showed an increase maturation index over time (49.18 ± 1.63% 76.63 1.22%, at 9 11 days, respectively) similar force contraction mono- (43.4 2.28 µN) co-cultured (47.6 4.7 tissues. Four weeks after implantation subcutaneous pockets nude rats, vessel length density within constructs significantly higher SVF (5.03 0.29 mm/mm2) compared monocultured (3.68 0.32 (p < 0.005). Although no mature neuromuscular junctions present, nerve-like structures predominantly observed cells. demonstrates that cells can support both vivo vascularization innervation tissues, making significant progress towards clinical translation.