Corneal Regeneration Methods and Protocols

This volume details different laboratory techniques used to develop regenerative therapies that help treat corneal blindness. Chapters guide readers through methods and protocols on optimal cell culture conditions, gene-editing techniques, multiple types of scaffold for corneal regeneration, how to evaluate the success of these therapies, and cell and material characterization techniques. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Isolation and Culture of Corneal Stromal Stem Cells.- In vitro Expansion of Corneal Endothelial Cells for Transplantation.- Primary Culture of Cornea-Limbal Epithelial Cells In Vitro.- Optimization of Human Limbal Stem Cell Culture by Replating a Single Limbal Explant.- A Guide to the Development of Human Cornea Organoids from Induced Pluripotent Stem Cells in Culture.- Gene Editing for Corneal Stromal Regeneration.- Preparation and Administration of Adeno-associated Virus Vectors for Corneal Gene Delivery.-The Self-assembly Approach as a Tool for the Tissue Engineering of a Bi-lamellar Human Cornea.- Formation of Corneal Stromal-like Assemblies using Human Corneal Fibroblasts and Macromolecular Crowding.- Preparation of Dried Amniotic Membrane for Corneal Repair.- Fabrication of Corneal Extracellular Matrix Derived Hydrogels.- Synthesis and Application of Collagens for Assembling a Corneal Implant.- Development and Validation of a 3D in vitro Model to Study the Chemotactic Behaviour of Corneal Stromal Fibroblasts.- Femtosecond Laser Assisted Surgery for Implantation of Bioengineered Corneal Stroma to Promote Corneal Regeneration.- The use of Animal Models to Assess Engineered Corneal Tissue.- X-ray Diffraction Imaging of Corneal Ultrastructure.