Article Figures & Data
Figures
- Fig. 1.
Schematic illustration of six corneal models using chambers with thin and thick CVM scaffolds. BM model (A), CEpi model (B), CEpi-Endo model (C), AS model (D), CEpi-AS model (E), and CEpi-AS-Endo model (F). Black lines represent plastic cylinders and hangers of a CVM chamber.
- Fig. 2.
Phase-contrast microscopic observations of BCD C/D-1b cells. BCD C/D-1b cells before cloning (A) and endothelial cell–like clonal cells at passage numbers 1 (B) and 28 (C) after cloning were cultured in a tissue culture plate. Scale bar, 50 μm.
- Fig. 3.
Microscopic observations of a corneal endothelial layer in a CVM camber. The endothelial cell–like clonal cells from BCD C/D-1b cells were cultured for 1 day (A, D, and G), 3 days (B, E, and H), and 7 days (C, F, and I). The clonal cells were observed with a phase-contrast microscope (A–C), and were stained with antibodies for Na+-K+ ATPase (D–F) and ZO-1 (G–I). The nuclei of cells were counterstained with Hoechst 33342 (D–I). Scale bar, 50 μm.
- Fig. 4.
Time-dependent change of TEER-end values of a corneal endothelial layer in a CVM camber. Each value represents the mean ± S.D. (n = 3). *P < 0.05.
- Fig. 5.
Microscopic observations of a CEpi-AS model. Cross sections of the model were stained with hematoxylin and eosin (A and B) and were also stained with antibodies for ZO-1 (C), occludin (D), connexin-43 (E), cytokeratin 3 (F), and MUC1 (G). The nuclei of cells were counterstained with Hoechst 33342 (C–G). Scale bar, 50 μm.
- Fig. 6.
Microscopic observations of corneal models composed of corneal epithelial cells and endothelial cells via a CVM. Cross sections of the CEpi-Endo model (A) and the CEpi-AS-Endo model (B) were stained with hematoxylin and eosin. Scale bar, 50 μm.
- Fig. 7.
Comparison among the permeability coefficient of a BM model (A), that of an AS model (B), and that of a commercially available multiporous PET membrane (C). Each permeability coefficient was tested using cyanocobalamin, FD4, FD10, FD20, and FD40. Dashed lines represent slopes calculated by least-squares test. Each value represents the mean ± S.D. (n = 3).
- Fig. 8.
Comparison between the permeability coefficient of FD-4 (A) and FD-10 (B) in each corneal model reconstructed in the CVM chamber described in Table 1 and that in an excised rabbit cornea. Each value represents the mean ± S.D. (n = 3). *P < 0.1; **P < 0.05; ***P < 0.01; ****P < 0.001.
Tables
- TABLE 1
Permeability coefficient of six corneal models
Each value represents the mean ± S.D. of three independent experiments.
Type of Model Permeability Coefficient Cyanocobalamin FD-4 FD-10 FD-20 FD-40 ×10−6 cm/s Acellular model BM 9.38 ± 0.47 4.22 ± 0.18 1.20 ± 0.31 0.46 ± 0.06 0.12 ± 0.002 AS 4.45 ± 0.32 1.46 ± 0.14 0.81 ± 0.11 0.24 ± 0.03 0.17 ± 0.03 Culture model CEpi 0.82 ± 0.097 0.36 ± 0.01 0.31 ± 0.02 0.36 ± 0.06 0.054 ± 0.01 CEpi-Endo Not detected 0.25 ± 0.04 0.13 ± 0.02 0.084 ± 0.01 0.050 ± 0.01 CEpi-AS Not detected 0.14 ± 0.06 0.046 ± 0.07 0.009 ± 0.002 0.017 ± 0.009 CEpi-AS -Endo Not detected 0.033 ± 0.02 0.019 ± 0.01 0.016 ± 0.005 0.033 ± 0.028
