Main 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.

Contents:

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 Raeder

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