Corneal extracellular matrix decellularization

Abstract

Decellularized corneal scaffolds have the potential to be used as alternatives to donor corneas during keratoplasty. Here a decellularization technique is described that involves the use of sodium dodecyl sulfate, Triton-X100, DNAse and RNAse to remove cells and cellular constituents. We have previously found that this combination of chemicals and enzymes to be effective at removing cells while retaining extracellular matrix proteins. In addition, different methods for assessing if the decellularization process has been successful are discussed. These include techniques to identify and quantify the presence of cells, DNA and extracellular matrix components as well as methods to examine the collagen fibril organization and scaffold transparency.

Keywords: Collagen; Cornea; Extracellular matrix; Keratocyte; Scaffold; Stroma.