Fig. 6

Bioengineered multi-lamellar human corneal stroma tissue constructs in vitro. a Schematic pictures of the assembly process for three-dimensional corneal stroma tissue constructs. Six patterned silk membranes with seeded PDLSCs (orange cells) were stacked one by one in a 24-well plate, with two plastic rings on the top of the construct (b) to keep the membranes close together. The constructs were cultured in keratocyte differentiation medium with 1 μM substance P (green lines in a) for 18 days. c Macroscopic picture shows that the constructs were clear enough to allow visualization of the letters below after 18 days of culture. d H&E staining showed the cells growing between every two layers (d1–d3). e–g Representative pictures show the fine integration of cells and silk membranes. h The expression of collagens and proteoglycans in bioengineered human corneal stroma was evaluated by immunofluorescence staining on frozen sections. The right panels of the bioengineered human corneal stroma images show the merged picture of the left panels (ECM component staining) and the middle panels (DAPI staining). The far right panels show the negative control (NTC) without primary antibody. S silk membrane