Organ Regeneration Based on Developmental Biology

143

a

b

Harderiangland germ

Bioengineered

Day 0 Day 3 Day 5

Lacrimalgland germ

In vitro culture

Day

30

after

transplantation

c

Duct association

d

NF-H/calponin

AQP5/

E-cadherin

Calponin/

H & E staining E-cadherin

Bioengineered

lacrimal gland

Bioengineeredlacrimal glan

d

Bioengineered

harderian gland

Fig. 8.3 Generation of bioengineered lacrimal gland germ using the organ germ method. Phase-
contrast images of development of the bioengineered lacrimal gland germ (upper line) and bioen-
gineered harderian gland germ (lower line). Scale bar, 100 μm. Photographs of the bioengineered
lacrimal gland germ after transplantation. At 30 days after transplantation, the bioengineered lac-
rimal gland and bioengineered harderian gland were engrafted and developed (right and left; scale
bar, 500 μm). Histological analysis of the duct association between the bioengineered lacrimal
gland and recipient lacrimal excretory duct. Bioengineered lacrimal glands regenerated using
DsRed transgenic mouse-derived epithelial cells (red), duct association between the bioengineered
lacrimal gland and the recipient mouse (arrowhead). FITC-gelatin (green) which was injected
from the recipient lacrimal excretory duct could successfully reach to the bioengineered lacrimal
gland (yellow). DAPI (blue) and the excretory duct (dotted line) are shown. Scale bars, 100 μm.
Histological and immunohistochemical analysis of the bioengineered lacrimal gland after trans-
plantation. The HE staining revealed that the bioengineered lacrimal gland achieved a mature
secretory gland structure including acini and duct (left; scale bar, 50 μm). Aquaporin-5 is red and
E-cadherin is green in the center-left panel. Calponin is red and E-cadherin is green in the center-
right panel. Calponin is red, neurofilament-H (NF-H) is green, and DAPI is blue in the right panel.
Scale bars, 50 μm (Modified and reprinted from Hirayama et al.^112 )

8 Functional Lacrimal Gland Regeneration