Fig. 4: Preparation of muscle and fat mixed cultured meat.

A Experimental design for porcine fibroblast myogenic and lipogenic differentiation in 3D culture. B Representative images of MHC and Oil Red O staining of cells upon myogenesis/lipogenesis in 3D culture. Scale bars, 50 µm. C Fluorescence imaging of cell viability assay of three-dimensional GelMA constructs under differentiation conditions. Live/dead staining with calcein-AM (green, live cells) and propidium iodide (red, dead cells), with nuclei counterstained with Hochest (blue). Scale bars, 50 µm. D, E Expression of myogenic genes and lipogenic genes in the cells of cultured meat and 3D porcine fibroblasts without any stimulation(control) by RT-qPCR. Error bars indicate s.e.m (n = 3 independent cultures). *P < 0.05, **P < 0.01, ***P < 0.001. ****P < 0.0001. F Western blotting showing the expression levels of MHC and GAPDH in porcine fibroblasts undifferentiation(control), myogenic differentiation, myogenic and lipogenic differentiation in 3D culture. n = 3. G Quantification of relative protein expression normalized to GAPDH. Band intensities were measured using ImageJ, and values represent the ratio of target protein to GAPDH from three independent experiments. Data are shown as mean ± SD. Statistical significance was determined using one-way ANOVA. *p < 0.05 was considered significant. H Macroscopic morphology of the GelMA hydrogel scaffold(left) and cultured meat (right). Scale bars, 1 cm. I Microstructure of 3D GelMA constructs under different differentiation conditions by scanning electron microscopy. Scale bars, 100 µm.