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. Authoritative and cutting-edge, Corneal Regeneration: Methods and Protocols aims to be useful for new and experienced laboratory researchers working on different aspects of corneal regeneration.

Isolation and Culture of Corneal Stromal Stem Cells / Richard M. Nagymihaly, Morten C. Moe, Goran Petrovski
In Vitro Expansion of Corneal Endothelial Cells for Transplantation / Kim Santerre, Isabelle Xu, Mathieu Thériault, Stéphanie Proulx
Primary Culture of Cornea-Limbal Epithelial Cells In Vitro / Finbarr O’Sullivan
Optimization of Human Limbal Stem Cell Culture by Replating a Single Limbal Explant / Marina López-Paniagua, Teresa Nieto-Miguel, Sara Galindo, Laura García-Posadas, Ana de la Mata, Rosa M. Corrales et al.
A Guide to the Development of Human CorneaOrganoids from Induced Pluripotent Stem Cells in Culture / James W. Foster, Karl J. Wahlin, Shukti Chakravarti
Gene Editing for Corneal Stromal Regeneration / Tara Moore, Connie Chao-Shern, Larry DeDionisio, Kathleen A. Christie, M. Andrew Nesbit
Preparation and Administration of Adeno-associated Virus Vectors for Corneal Gene Delivery / Liujiang Song, Jacquelyn J. Bower, Matthew L. Hirsch
The Self-assembly Approach as a Tool for the Tissue Engineering of a Bi-lamellar Human Cornea / Gaëtan Le-Bel, Pascale Desjardins, Camille Couture, Lucie Germain, Sylvain L. Guérin
Formation of Corneal Stromal-Like Assemblies Using Human Corneal Fibroblasts and Macromolecular Crowding / Mehmet Gürdal, Gülinnaz Ercan, Dimitrios I. Zeugolis
Preparation of Dried Amniotic Membrane for Corneal Repair / Andrew Hopkinson, Emily R. Britchford, Laura E. Sidney
Fabrication of Corneal Extracellular Matrix-Derived Hydrogels / Mark Ahearne, Julia Fernández-Pérez
Synthesis and Application of Collagens for Assembling a Corneal Implant / Elle Edin, Fiona Simpson, May Griffith
Development and Validation of a 3D In Vitro Model to Study the Chemotactic Behavior of Corneal Stromal Fibroblasts / Evrim Ceren Kabak, Julia Fernández-Pérez, Mark Ahearne
Femtosecond Laser-Assisted Surgery for Implantation of Bioengineered Corneal Stroma to Promote Corneal Regeneration / Neil Lagali, Mehrdad Rafat
The Use of Animal Models to Assess Engineered Corneal Tissue / Robert Thomas Brady, Peter W. Madden
X-Ray Diffraction Imaging of Corneal Ultrastructure / Keith M. Meek, Andrew J. Quantock, Sally Hayes, James Bell