Description
Continuous regeneration of the cornea is necessary to maintain this tissue in the transparent state that is essential for vision. Therapy for repair of the damaged anterior cornea is currently addressed through the transplantation of donor corneas or the delivery of limbal epithelial stem cells (LESC) to the ocular surface using amniotic membrane (AM) as a supporting scaffold. Research on the bioengineering of corneal equivalents as replacement tissue is underway to develop viable corneal prosthetics. Corneal Regenerative Medicine: Methods and Protocols provides a concise overview of essential techniques in the field of corneal regenerative medicine, highlighting novel strategies to guide the management of key therapies within this area of medicine. Divided into four convenient sections, topics include the identification, characterisation and cultivation of LESC, as well as the investigation of biopolymers used as the basis for corneal substitutes. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, Corneal Regenerative Medicine: Methods and Protocols covers the fundamental techniques useful for both the laboratory and clinical settings. Part I. Perspectives 1. Limbal Epithelial Cell Therapy: Past, Present and Future Tor Paaske Utheim 2. The Artificial Cornea Naresh Polisetti, Mohammad Mirazul Islam, and May Griffith Part II. Corneal Stem Cell Enrichment and Characterisation 3. Clonal Analysis of Limbal Epithelial Stem Cell Populations Ursula Schlootzer-Schrehardt 4. The Use of Bromodeoxyuridine Incorporation Assays to Assess Corneal Stem Cell Proliferation Ashley M. Crane and Sanjoy K. Bhattacharya 5. Enrichment of Human Corneal Epithelial Stem/Progenitor Cells by Magnetic Bead Sorting Using SSEA4 as a Negative Marker Martin N. Nakatsu and Sophie X. Deng 6. Limbal Epithelial Stem Cell Identification using Immunoblotting Analysis Bernice Wright and Che J. Connon Part III. Corneal Cell Culture 7. The Culture of Limbal Epithelial Cells Tor Paaske Utheim, Torstein Lyberg, and Sten Rder 8. The Culture of Limbal Stromal Cells and Corneal Endothelial Cells Naresh Polisetti and Nancy C. Joyce Part IV. Development of the Artificial Cornea 9. The Formation of a Novel Tissue-Engineered Cornea using Collagen Scaffolds and Limbal Stem Cells Shengli Mi and Che J. Connon 10. Fabrication of a Human Recombinant Collagen-based Corneal Substitute using Carbodiimide Chemistry Mohammad Mirazul Islam, May Griffith, and Kimberley Merrett 11. Fabrication of a Corneal-Limbal Tissue Substitute Using Silk Fibroin Laura J. Bray, Karina A. George, Shuko Suzuki, Traian V. Chirila, and Damien G. Harkin 12. Cultivation of Limbal Epithelial Cells on Electrospun Poly (lactide-co-glycolide) Scaffolds for Delivery to the Cornea Pallavi Deshpande, Charanya Ramachandran, Virender S. Sangwan, and Sheila MacNeil 13. The Growth and Delivery of Mesenchymal and Limbal Stem Cells using Copolymer Polyamide 6/12 Nanofiber Scaffolds Vladimir Holan, Eliska Svobodova, and Peter Trosan 14. Encapsulation and Culture of Mammalian Cells including Corneal Cells in Alginate Hydrogels Nicola C. Hunt and Liam M. Grover 15. In vivo Confocal Microscopy of the Cornea to Assess Tissue Regenerative Response after Biomaterial Implantation in Humans Neil Lagali, May Griffith, and Per Fagerhorn
