Successful Transplantation of In Vitro Expanded Human Cadaver Corneal Endothelial Precursor Cells On to a Cadaver Bovine’s Eye Using a Nanocomposite Gel Sheet

Abstract

Purpose: In vitro expansion of human corneal endothelial precursor (HCEP) cells has been reported via production of cell aggregated spheres. However, to translate this procedure in human patients warrants maintaining the position of the eyeballs facing down for 36 h, which is not feasible. In this study, we report a method using a nanocomposite (NC) gel sheet to accomplish the integration of HCEP cells to the endothelium of cadaver bovine’s eyes.

Materials and Methods: HCEP cells were isolated from the corneal endothelium of a cadaver human eye and then expanded using a thermoreversible gelation polymer (TGP) as reported earlier. For the study, three cadaver bovine eyes were used. The NC gel sheets were inserted into the bovine eyes’, aligned and suture-fixed in position under the host endothelium. HCEP cells previously expanded in the TGP were harvested and injected using a 26-gauge syringe between the endothelium and the NC gel sheet. The eyes were left undisturbed for three hours following which the NC gel sheets were gently removed. The corneas were harvested and subjected to histopathological studies.

Results: Histopathological studies showed that all the three corneas used for NC gel sheet implantation showed the presence of engrafted HCEP cells, seen as multi-layered cells over the native endothelium of the bovine cornea. Examination of the NC gel sheets used for implantation showed that only very few corneal endothelial cells remained on the sheets amounting to what could be considered negligible.

Conclusion: The use of the NC gel sheet makes HCEP cell transplantation feasible for human patients. Further in vitro basic studies followed by translational studies are necessary to bring this method for clinical application in appropriate indications.

• Bovine cornea
• corneal endothelial diseases
• corneal endothelial precursor cell transplantation
• human corneal endothelial precursor (HCEP) cells
• nanocomposite (NC) gel sheet

Acknowledgements

The authors acknowledge

1. Prof. Masaru Iwasaki, Department of Clinical Research, Yamanashi University, Japan, and M/S Chennai Cell Cluster (CCC) for technical advice.

2. Loyola ICAM College of Engineering Technology (LICET) and Loyola Institute of Frontier Energy (LIFE) for their support to our research work.

3. Dr. Senthilkumar Preethy for her assistance in preparation of the manuscript.