Corneal Regenerative Medicine, 2013 Methods and Protocols Methods in Molecular Biology Series, Vol. 1014

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.  

Limbal Epithelial Cell Therapy: Past, Present and Future.- The Artificial Cornea.- Clonal Analysis of Limbal Epithelial Stem Cell Populations.- The Use of Bromodeoxyuridine Incorporation Assays to Assess Corneal Stem Cell Proliferation.- Enrichment of Human Corneal Epithelial Stem/Progenitor Cells by Magnetic Bead Sorting Using SSEA4 as a Negative Marker.- Limbal Epithelial Stem Cell Identification using Immunoblotting Analysis.- The Culture of Limbal Epithelial Cells.- The Culture of Limbal Stromal Cells and Corneal Endothelial Cells.- The Formation of a Novel Tissue-Engineered Cornea using Collagen Scaffolds and Limbal Stem Cells.- Fabrication of a Human Recombinant Collagen-based Corneal Substitute using Carbodiimide Chemistry.- Fabrication of a Corneal-Limbal Tissue Substitute Using Silk Fibroin.- Cultivation of Limbal Epithelial Cells on Electrospun Poly (lactide-co-glycolide) Scaffolds for Delivery to the Cornea.- The Growth and Delivery of Mesenchymal and Limbal Stem Cells using Copolymer Polyamide 6/12 Nanofiber Scaffolds.- Encapsulation and Culture of Mammalian Cells including Corneal Cells in Alginate Hydrogels.- In vivo Confocal Microscopy of the Cornea to Assess Tissue Regenerative Response after Biomaterial Implantation in Humans.