Fig. 5: pgEpiSCs-derived myoblast attachment and maturation on plant-based 3D scaffolds of Ca2+-KGM5-SA5.
From: Generation of three-dimensional meat-like tissue from stable pig epiblast stem cells
a Schematic diagram of the inoculation of pgEpiSCs differentiated to stage V and subsequently inoculated on plant-based 3D edible scaffolds, proliferated for 5 days, and cultured in N2 serum-free terminal differentiation medium for 5 days. pgEpiSCs-CM: pgEpiSCs-derived CM. b The cells were stained with Calcein-AM (green) and PI (red) after inoculation into the scaffold culture for 24 h. Scale bar, 100 μm. c SEM images of pgEpiSCs-derived CM after culture. Scale bar, 100 μm (left) and 50 μm (right). d Confocal microscopy to observe the differentiation status on plant-based 3D edible scaffolds, representative fluorescent images of cytoskeletal protein F-actin in day 10. Red, phalloidin; blue, DAPI. Scale bar, 50 μm. e Quantification of mRNA expression of pluripotency (OCT4, NANOG), maturation of myogenic differentiation (MYOG, MYMK), skeletal muscle fibers (MYH2, MYH3) and collagen formation (COL3A1, COL5A2) related genes by qRT-PCR. WT: undifferentiated pgEpiSCs; 3D differentiation: pgEpiSCs-MCs differentiation on 3D scaffolds. f Appearance of stained pgEpiSCs-derived CM in culture for 10d. g Textural properties of pgEpiSCs-derived CM in culture for 10d. The red line indicates the value of the fresh pork. Blank: empty scaffolds without cell inoculation; pg-CM: pgEpiSCs-derived CM. For (e, g), error bars indicate means ± SD, n = 3. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, and ns indicates p > 0.05. Represent significant using two-tailed student’s t test and similar results were obtained in three independent experiments. Exact P values are listed in Source Data Fig. 5.
