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Jui-Yang Lai, PhD; Ko-Hua Chen, MD; Wen-Ming Hsu, MD; Ging-Ho Hsiue, PhD; Yen-Hsien Lee, MS

Arch Ophthalmol. 2006;124:1441-1448.

Objective  To investigate whether the bioengineered human corneal endothelial cell (HCEC) monolayers harvested from thermoresponsive culture supports could be used as biological tissue equivalents.

Methods  Untransformed adult HCECs derived from eye bank corneas were cultivated on a thermoresponsive poly-N-isopropylacrylamide–grafted surface for 3 weeks at 37°C. Confluent cell cultures with a phenotype and cell density similar to HCECs in vivo were detached as a laminated sheet by lowering the culture temperature to 20°C. In vitro characteristics of the HCEC sheets were determined evaluating their viability and by scanning electron microscopy, immunohistochemistry, and histological studies.

Results  After separation from culture surfaces via a thermal stimulus, the HCEC sheets remained viable. Polygonal cell morphology and multiple cellular interconnections were observed throughout the HCEC sheets. Immunolocalization of zonula occludens-1 and Na⁺,K⁺–adenosine triphosphatase (ATPase) indicated the formation of tight junctions and the distribution of ionic pumps at the cell boundary. In addition, we ascertained that cultured HCECs have a monolayered architecture that mimics native corneal endothelium.

Conclusion  These data suggest that a well-organized and functional HCEC monolayer can feasibly be used as tissue equivalents for replacing compromised endothelium.

Clinical Relevance  Bioengineered human corneal endothelium fabricated from thermoresponsive supports can potentially offer a new therapeutic strategy for corneal endothelial cell loss.

Author Affiliations: Department of Chemical Engineering, National Tsing Hua University, Hsinchu, Taiwan (Drs Lai and Hsiue); and Division of Medical Engineering Research, National Health Research Institutes (Drs Lai and Chen and Mr Lee), and Department of Ophthalmology, Taipei Veterans General Hospital (Drs Chen and Hsu), Taipei, Taiwan.

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