Fig. 1

Immunofluorescence and cross-sectional H&E staining of porcine cell-seeded-RAFT (A) Schematic diagram showing that excessive hydrogel water was wicking into the absorber to produce RAFT. After that, PCECs were seeded on RAFT (0.4 ml of 5 mg/ml collagen I) coated with collagen IV and culture for 7 days to construct a porcine cell-seeded-RAFT. (B) The light transmission of RAFT across wavelengths from 400 to 700 nm, n = 3 biological replicates. (C) Camera image of RAFT in a 24-well plate. (D, H) Immunofluorescence staining results showed the expression of ZO-1, Na/K ATPase, N-Cadherin and F-actin on RAFT and control. The control was PCECs seeded on a round glass cover slip at the same seeding density as on the cell-seeded RAFT. Continuous expression of ZO-1 was observed at the cell–cell borders on RAFT and control. (E, I) The expression of Na/K ATP at the cell borders and cell membrane on RAFT and control. (F, J) The expression of N-Cadherin at the cell borders and cell membrane on RAFT and control. (G, K) The expression of F-actin filaments showed the polygonal shape of cells on RAFT and control. Images are representative of n = 3 biological replicates. Scale bar = 50 µm. (L) The cross-sectional H&E staining showed that RAFT was a compressed, homogenous, and dense collagen hydrogel. (M) PCECs formed a mono-cell layer on the surface of the cell-seeded-RAFT (➤). Images are representative of n = 3 biological replicates. Scale bar = 100 µm.