Journal of Clinical and Experimental Ophthalmology
Open Access
ISSN: 2155-9570
+44 1223 790975
ISSN: 2155-9570
+44 1223 790975
Tanvi Khanna
India
Research Article
Standardization of Human Corneal Endothelial Cell Isolation and the Use of Denuded Human Amniotic Membrane as a Scaffold for Human Corneal Endothelial cells
Author(s): Kalpana Suresh, Alan Mathew Punnoose, Sarah Kuruvilla and Tanvi KhannaKalpana Suresh, Alan Mathew Punnoose, Sarah Kuruvilla and Tanvi Khanna
Objectives: To standardize the isolation of human corneal endothelial cells (HCECs) and to use the denuded human amniotic membrane (HAM) as a scaffold for isolated HCECs.
Methods: Human amniotic membrane was denuded using 1.2 units/ml of Dispase II at 37°C for 60 minutes followed by mechanical scraping. Corneal endothelial and Descemet’s membrane sheets were peeled from human donor cadaveric eyes unfit for surgical use and enzymatically digested with 2 mg/ml of collagenase II solution at 37°C and 5% CO2 for 2 hrs. Isolated cells were resuspended in culture medium with supplements and plated onto uncoated cultureware for four hours to eliminate fibroblasts which adhere more rapidly than endothelial cells. After preplating, the non-adherent cells were seeded onto gelatin coated dishes or onto denuded amniotic membrane in OptiMEM med.. View More»
DOI:
10.4172/2155-9570.1000421