title: Characterization of the limbal epithelial stem cell niche: novel imaging techniques permit in vivo observation and targeted biopsy of limbal epithelial stem cells.
abstract: It is anticipated that stem cell (SC) therapy will enable the regeneration of diseased tissues and organs. Understanding SC niches is an essential step toward realizing this goal. By virtue of its optical transparency and physical separation of SC and transient amplifying cell compartments, the human cornea provides a unique opportunity to visualize and observe a population of adult stem cells, limbal epithelial stem cells (LESCs), in their niche environment. To date, the characteristics of the LESC niche have remained unclear. State-of-the-art imaging techniques were used to construct a three-dimensional (3D) view of the entire human corneal limbus and identify the structural characteristics of the LESC niche. Two distinct candidate LESC niche structures were identified. Cells within these structures express high levels of the putative limbal stem cell markers p63alpha and ABCG2; however, current methods cannot identify for certain which exact cells within this cell population are truly LESCs. These structures could be located and observed in vivo in normal human subjects, but not in patients with clinically diagnosed corneal LESC deficiency. The distribution of these structures around the corneal circumference is not uniform. Biopsies targeted to limbal regions rich in LESC niche structures yielded significantly higher numbers of LESCs in culture. Our findings demonstrate how adult stem cell niches can be identified and observed in vivo in humans and provide new biological insight into the importance of LESC niche structures in maintaining normal LESC function. Finally, the concept of targeted biopsy of adult SC niches improves stem cell yield and may prove to be essential for the successful development of novel adult stem cell therapies. Disclosure of potential conflicts of interest is found at the end of this article.
authors.affiliation: MRC Clinical Research Fellow, Institute of Ophthalmology and Moorfields Eye Hospital, 11-43 Bath Street, London, EC1V 9EL, United Kingdom. a.shortt@ucl.ac.uk
authors.forename: Alex J
authors.initials: AJ
authors.lastname: Shortt
authors.forename: Genevieve A
authors.initials: GA
authors.lastname: Secker
authors.forename: Peter M
authors.initials: PM
authors.lastname: Munro
authors.forename: Peng T
authors.initials: PT
authors.lastname: Khaw
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Tuft
authors.forename: Julie T
authors.initials: JT
authors.lastname: Daniels
mesh_terms: D000293:Adolescent; D000328:Adult; D053687:Adult Stem Cells / Q000166:cytology* / Q000648:ultrastructure; D000368:Aged; D000369:Aged, 80 and over; D001706:Biopsy / Q000379:methods*; D048429:Cell Size; D002478:Cells, Cultured; D002648:Child; D002675:Child, Preschool; D003952:Diagnostic Imaging; D019573:Epithelium, Corneal / Q000166:cytology* / Q000473:pathology; D005260:Female; D006801:Humans; D016850:Limbus Corneae / Q000166:cytology* / Q000473:pathology / Q000648:ultrastructure; D008297:Male; D008875:Middle Aged; D017154:Stromal Cells / Q000166:cytology