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Biomedical Sciences - Molecular Medicine | Corneal Regenerative Medicine - Methods and Protocols

Corneal Regenerative Medicine

Methods and Protocols

Series: Methods in Molecular Biology, Vol. 1014

Wright, Bernice, Connon, Che J (Eds.)

2013, XII, 230 p. 56 illus., 33 illus. in color.

A product of Humana Press
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  • Includes cutting-edge methods and protocols
  • Provides step-by-step detail essential for reproducible results
  • Contains key notes and implementation advice from the experts

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.  

Content Level » Professional/practitioner

Keywords » artificial cornea - corneal cell structure - corneal stem cell - corneal substitute - limbal epithelial cell - limbal stem cell

Related subjects » Molecular Medicine - Ophthalmology

Table of 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 Ræder


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

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