ReLEx SMILE Method to Expansion and Create Scaffolds from Corneal Fibroblasts

To identify three-dimensional corneal stromal constructs that could promote a human corneal fibroblasts survival. Small-incision lenticule extraction was used for obtained corneal stroma for in vitro expansion of primary human corneal fibroblasts; monoclonal antibodies to keratocyte, mesenchymal stem cell/fibroblast and epithelial cell was used to phenotyping expanded human corneal fibroblast on the flow cytometry; polyethylene glycol, platelet lysate and methylene cellulose was used to prepare three-dimensional corneal stroma constructs, morphology was monitored by phase contrast microscopy. For 3-4 weeks human corneal fibroblasts were expanded in vitro from lenticule. Obtained cells expressed mesenchymal stem cell/fibroblast surface marker and was negative for keratocyte and epithelial cells surface markers. A combination of polyethylene glycol with platelet lysate significantly increased growth and survival of human corneal fibroblasts in three-dimensional corneal stroma construct.