Physiomimetic 3D skeletal muscle reveals senescence-driven matreotype remodeling driving paracrine senescence and regenerative failure

We developed a physiomimetic, 3D engineered skeletal muscle platform by embedding C2C12 myoblasts within muscle tissue–derived extracellular matrix (mdECM) to model how cellular senescence remodels the muscle niche and propagates dysfunction. Senescence was induced in 3D constructs by a single 20 Gy X-ray irradiation, resulting in impaired myogenic differentiation and structural maturation. To define senescence-associated transcriptional programs and paracrine senescence, we performed bulk RNA-sequencing on (i) irradiated senescent (SnC) versus (ii) non-irradiated normal 3D muscle constructs, and (iii) normal constructs exposed in a co-culture to senescent construct–derived secretome (SASP). These datasets enable quantitative characterization of senescence-driven gene expression changes linked to ECM remodeling, inflammation, and loss of myogenic/contractile programs in a physiologically relevant 3D muscle model.