Figure 3. Histological analysis of a bioengineered gland.

(a) Immunohistochemical analysis of the acinar and duct cells in natural or bioengineered lacrimal and harderian glands. Their respective acini were analysed by immunostaining with specific antibodies for AQP5(red, in left panels), calponin (red, in centre and left panels), neurofilament-H (NF-H, green, in right panels) and E-cadherin (green, in left and centre panels). Nuclei were stained using Hoechst 33342 dye (blue). Scale bar, 50 μm. (b) Immunohistochemical analysis of a bioengineered lacrimal gland reconstituted between H2B-GFP-transgenic mice-derived epithelial cells and normal mice-derived mesenchymal cells. Green fluorescence indicates the nuclei of epithelial cells isolated from H2B-GFP-transgenic mice. The myoepithelial cells were analysed by immunostaining with specific antibodies for calponin (red). The boxed area in the left panel is shown at a higher magnification in the right panel. Scale bar, 50 μm in the upper panel, 20 μm in the lower panel. (c) Photomicrographs showing lactoferrin (left) in the acini of a natural lacrimal gland (upper) and a bioengineered lacrimal gland (lower). Lipids (right) in the natural harderian gland (upper) and bioengineered harderian gland (lower) acini were revealed by oil-red O staining. Scale bar, 50 μm.