Properties of corneas reconstructed with cultured human corneal endothelial cells and human corneal stroma.
OBJECTIVE
To examine the properties of corneas tissue-engineered with cultured human corneal endothelial cells (HCEC) and human corneal stroma.


METHODS
Primary HCEC cultures were established from endothelial cell layer explants and propagated on culture dishes coated with bovine corneal endothelial extracellular matrix. A cell suspension of HCEC at the fifth passage was transferred onto human corneal stroma deprived of endothelial cells, and the corneas were gently centrifuged to enhance cell attachment. The cell density of the tissue-engineered corneas was examined after staining with alizarin red and trypan blue. The tissue-engineered corneas were histologically examined by light and electron microscopy. The pump function of the tissue-engineered corneas was measured using an Ussing chamber.


RESULTS
The mean endothelial cell density of four tissue-engineered corneas was 2380 +/- 264 cells/mm(2) (mean +/- SD). HCEC on the tissue-engineered corneas had a morphology similar to HCEC in vivo. The pump function parameters of the tissue-engineered corneas were 55%-75% of those of normal corneas.


CONCLUSIONS
HCEC on the tissue-engineered corneas have morphology and cellular density similar to HCEC in vivo, whereas the pump function of the tissue-engineered corneas was lower than in normal corneas.
Amano
Shiro
S
Department of Ophthalmology, University of Tokyo School of Medicine, Tokyo, Japan. amanos-tky@umin.ac.jp.
Mimura
Tatsuya
T
Department of Ophthalmology, University of Tokyo School of Medicine, Tokyo, Japan.
Yamagami
Satoru
S
Department of Ophthalmology, University of Tokyo School of Medicine, Tokyo, Japan.
Osakabe
Yasuhiro
Y
Meiwakai Medical Foundation, Miyata Eye Hospital, Miyakonojo, Miyazaki, Japan.
Miyata
Kazunori
K
Meiwakai Medical Foundation, Miyata Eye Hospital, Miyakonojo, Miyazaki, Japan.
D000328:Adult; D000368:Aged; D002452:Cell Count; D002478:Cells, Cultured; D003315:Cornea / Q000166:cytology* / Q000502:physiology; D003319:Corneal Stroma / Q000166:cytology*; D004728:Endothelium, Corneal / Q000166:cytology*; D006801:Humans; D008875:Middle Aged; D010539:Permeability; D023822:Tissue Engineering / Q000379:methods*