Fig. 6
Cell-laden multicomponent scaffolds. a Schematic of cell encapsulation within the hydrogel coating of the multicomponent devices. b 475 μm Z-stack of a single, excised window 24 h after MRC-5 lung fibroblasts were encapsulated in 7.5 wt% methacrylated gelatin hydrogel coating (1 × 106 cells mL−1). Cell nucleus was labeled with NucBlue (blue) and actin cytoskeleton was labeled with Alexa FluorTM 488 Phalloidin (green). c MRC-5 cells 1 day after encapsulation in 7.5 wt% methacrylated gelatin hydrogel coating (1 × 106 cells mL−1). Cell nucleus was labeled with NucBlue (blue) and actin cytoskeleton was labeled with Alexa FluorTM 488 Phalloidin (green). Maximum intensity projection of 100 μm Z-stack. d MRC-5 cells spread in the gel 14 days after encapsulation. e Percentage of cells that displayed functional esterase activity as measured by live/dead assay at 1, 14, and 28 days following MRC-5 encapsulation in 7.5 wt% methacrylated gelatin coatings. Values are displayed as mean + s.d. (n = 3). f Live/dead staining of NHDF-laden hydrogel 1 day after encapsulation in 7.5 wt% methacrylated gelatin hydrogel coating at a high density (10 × 106 cells mL−1). 270 μm Z-stack of a single, excised window. g NHDF-laden hydrogel 1 day after encapsulation in 7.5 wt% methacrylated gelatin hydrogel at a high density (10 × 106 cells mL−1). Cell nucleus was labeled with NucBlue (blue) and actin cytoskeleton was labeled with Alexa FluorTM 488 Phalloidin (green). Lateral view of a 388 μm Z-stack of a single, excised window. h Front view. i Schematic of co-culture scaffolds. j A monolayer of human mesenchymal stem cells (MSC) was seeded on the luminal surface of the tubular scaffold. Cell nucleus was labeled with NucBlue (blue), actin cytoskeleton was labeled with Alexa FluorTM 488 Phalloidin (green), and cell cytoplasm was labeled with CellTracker Red (red). k NHDF were encapsulated within a gel and GFP-expressing HUVEC were seeded on the luminal surface of the scaffold. Cell nucleus was labeled with NucBlue (blue), actin cytoskeleton was labeled with Alexa FluorTM 488 Phalloidin (green), and NHDF cell cytoplasm was labeled with CellTracker Red (red). 3D rendering of a 306 μm Z-stack of a single, excised window. l Lateral view of the co-culture scaffold. Dashed line indicates HUVEC layer on the luminal surface. Scale bars, 100 μm
