Optimization of Cultured Human Corneal Endothelial Cell Sheet Transplantation and Post-Operative Sheet Evaluation in a Rabbit Model

ABSTRACT

Purpose: To optimize cultured human corneal endothelial cell (cHCEC) sheet transplantation technique for maintenance of cHCEC viability.

Materials and Methods: cHCEC sheets cultured on a collagen scaffold were covered with or without Viscoat® and exposed to humidified air in the incubator. cHCEC sheets with or without Viscoat® were transplanted into cadaveric porcine eyes by the DSAEK technique with full air tamponade and incubated for various time periods. Then cell viability was determined by using the live/dead assay kit. cHCEC sheets with Viscoat® were transplanted into rabbit eyes and the sheets were histologically evaluated before and 14 days after transplantation.

Results: A collagen scaffold and Viscoat® were effective for protecting cHCEC from damage due to air exposure in vitro. All cells died after 18 hours of air exposure in porcine eyes in Viscoat® untreated control. In contrast, Viscoat® treatment sustained full cell viability following 2 hours and could maintain approximately 80% viability after 18 hours. In a rabbit model, transplanted cHCEC sheet with Viscoat® maintained cell density at 2803 ± 229 mm² (18% cell loss) and expression of N-cadherin, zonula occludens-1, and actin-filament localized to cell boundary as similar as donor HCEC.

Conclusion: Viscoat® can contribute to cHCEC protection from damage caused by exposure to air.

KEYWORDS:

• Air exposure
• collagen
• corneal endothelial cells
• regenerative medicine
• viscoat

Acknowledgments

The authors thank Ms Junko Nishida for excellent technical assistance.

Funding

Knowledge cluster initiative grant from the Japanese Ministry of Education, Culture, Sports, Science and Technology (MEXT). There is no commercial relationship in the form of financial support or personal financial interest.

Declaration of interest

The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the article.

Additional information

Funding

Knowledge cluster initiative grant from the Japanese Ministry of Education, Culture, Sports, Science and Technology (MEXT). There is no commercial relationship in the form of financial support or personal financial interest.