Fig. 3.

Fig. 3. Spinach Leaf Vascular Scaffolds in 3D Mammalian Cell Culture. Spinach leaf vascular scaffolds demonstrate remarkable retention of patency and perfusion capabilities post-decellularization, enabling successful adhesion and function of human pluripotent stem cell-derived cardiomyocytes (hPS-CMs) for 21 days. (A, B) Display the decellularized leaf before and after Ponceau Red perfusion, highlighting the preservation of the vascular architecture crucial for cellular perfusion and viability. (C, D) Fluorescence images depict leaf vasculature perfused with beads, illustrating the retention of 50 and 100 μm spheres within the vascular network. Scale bars indicate dimensions, crucial for assessing microvascular architecture: (C) 100 μm, (D) 500 μm. (E) hPS-CMs adhere to the leaf scaffold surface, forming distinct cell clusters critical for tissue organization and function. The scale bar denotes 50 μm, enabling precise assessment of cellular arrangements. (F) Contractile strain, indicative of cellular functionality, is visualized through a heatmap, providing insights into the dynamic behavior of hPSCMs within the scaffold environment. (G) Day 21 strain values reveal a diminished contractile strain magnitude, suggesting potential cellular maturation and adaptation within the scaffold over time. (H) Relative changes in fluorescent signals relative to the leaf surface are visualized, offering quantitative data on cellular distribution and viability throughout the scaffold. This comprehensive analysis demonstrates the efficacy of spinach leaf vascular scaffolds in supporting 3D mammalian cell culture, showcasing their potential in tissue engineering and regenerative medicine applications. The ability to retain vascular patency and perfusion, coupled with successful hPS-CM adhesion and function, underscores the utility of plant-derived scaffolds as biocompatible substrates for advanced cell culture studies. (Reproduced, with permission, copyright 2017, Elsevier)

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