title: Clonal analysis of limbal epithelial stem cell populations.
abstract: While convincing data clearly suggest the presence of stem cells in the basal limbal epithelium in vivo, testing the proliferation, self-renewal, and differentiation capacity of stem cells relies on the development of methodologies that allow for their isolation and extensive propagation in vitro. Clonal analysis involving differentiation between short-lived transient cell clones and long-lived stem cell clones is an invaluable technique to identify stem cells in vitro, and allows cells to be expanded over multiple passages. This chapter describes a protocol for the isolation, expansion, and clonal analysis of limbal epithelial stem cells. The cultivation method described may be essential for long-term restoration of the damaged ocular surface in patients with limbal stem cell deficiency.
authors.affiliation: Department of Ophthalmology, University of Erlangen-Nürnberg, Erlangen, Germany.
authors.forename: Ursula
authors.initials: U
authors.lastname: Schlötzer-Schrehardt
mesh_terms: D053687:Adult Stem Cells / Q000502:physiology* / Q000637:transplantation; D000818:Animals; D049109:Cell Proliferation; D018920:Coculture Techniques; D003316:Corneal Diseases / Q000628:therapy; D004847:Epithelial Cells / Q000502:physiology* / Q000637:transplantation; D019573:Epithelium, Corneal / Q000473:pathology; D061252:Feeder Cells; D006801:Humans; D016850:Limbus Corneae / Q000473:pathology; D051379:Mice; D041701:Swiss 3T3 Cells