Corneal Regeneration, 1st ed. 2020 Methods and Protocols Methods in Molecular Biology Series, Vol. 2145

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.

1. Isolation and Culture of Corneal Stromal Stem Cells

Richard M Nagymihaly, Morten C Moe, and Goran Petrovski

2. In vitro Expansion of Corneal Endothelial Cells for Transplantation

Kim Santerre, Isabelle Xu, Mathieu Thériault, and Stéphanie Proulx

3. Primary Culture of Cornea-Limbal Epithelial Cells In Vitro

Finbarr O'Sullivan 

4. Optimization of Human Limbal Stem Cell Culture by Replating a Single Limbal Explant

Marina López-Paniagua, Teresa Nieto-MiguelGalindo, Laura García-Posadas,Ana de la Mata, Rosa M. Corrales, Margarita Calonge,and Yolanda Diebold

5. A Guide to the Development of Human Cornea Organoids from Induced Pluripotent Stem Cells in Culture

James W Foster, Karl Wahlin, and Shukti Chakravarti

6. Gene Editing for Corneal Stromal Regeneration

Tara Moore, Connie Chao-Shern , Larry DeDionisio, Katie Christie, and Andrew M.Nesbit

7. Preparation and Administration of Adeno-associated Virus Vectors for Corneal Gene Delivery

Liujiang Song, Jacquelyn J. Bower, and Matthew L. Hirsch

8. 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, and Sylvain L. Guérin

9. Formation of Corneal Stromal-like Assemblies using Human Corneal Fibroblasts and Macromolecular Crowding

Mehmet Gürdal, Gülinnaz Ercan, and Dimitrios I. Zeugolis

10. Preparation of Dried Amniotic Membrane for Corneal Repair

Andrew Hopkinson , Emily R Britchford, and Laura E Sidney

11. Fabrication of Corneal Extracellular Matrix Derived Hydrogels

12. Synthesis and Application of Collagens for Assembling a Corneal Implant

Elle Edin, Fiona Simpson, and May Griffith

13. Development and Validation of a 3D in vitro Model to Study the Chemotactic Behaviour of Corneal Stromal Fibroblasts

14. Femtosecond Laser Assisted Surgery for Implantation of Bioengineered Corneal Stroma to Promote Corneal Regeneration

Neil Lagali and Mehrdad Rafat

15. The use of Animal Models to Assess Engineered Corneal Tissue

Robert Thomas Brady and Peter W Madden

Keith M Meek, Andrew J. Quantock, Sally Hayes, and James Bell

Includes cutting-edge methods and protocols

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts