Advanced imaging and tissue engineering of the human limbal epithelial stem cell niche.
The limbal epithelial stem cell niche provides a unique, physically protective environment in which limbal epithelial stem cells reside in close proximity with accessory cell types and their secreted factors. The use of advanced imaging techniques is described to visualize the niche in three dimensions in native human corneal tissue. In addition, a protocol is provided for the isolation and culture of three different cell types, including human limbal epithelial stem cells from the limbal niche of human donor tissue. Finally, the process of incorporating these cells within plastic compressed collagen constructs to form a tissue-engineered corneal limbus is described and how immunohistochemical techniques may be applied to characterize cell phenotype therein.
Department of Ocular Biology and Therapeutics, Institute of Ophthalmology, University College London, 11-43 Bath Street, London, EC1V 9EL, UK.
Isobel
I
Massie
Marc
M
Dziasko
Alvena
A
Kureshi
Hannah J
HJ
Levis
Louise
L
Morgan
Michael
M
Neale
Radhika
R
Sheth
Victoria E
VE
Tovell
Amanda J
AJ
Vernon
James L
JL
Funderburgh
Julie T
JT
Daniels
D018929:Cell Culture Techniques / Q000379:methods; D002469:Cell Separation / Q000379:methods; D019573:Epithelium, Corneal / Q000166:cytology*; D005347:Fibroblasts / Q000166:cytology; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D016850:Limbus Corneae / Q000166:cytology*; D018613:Microscopy, Confocal / Q000379:methods; D008855:Microscopy, Electron, Scanning / Q000379:methods; D055153:Stem Cell Niche; D013234:Stem Cells / Q000166:cytology*; D023822:Tissue Engineering / Q000379:methods*