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Sphere-forming corneal cells repopulate dystrophic keratoconic stroma:Implications for potential therapy

World Journal of Stem Cells 2020年第1期12卷 35-54页

作者： Himanshu Wadhwa Salim Ismail Jennifer J McGhee Bert Van der Werf trevor sherwin Department of Ophthalmology Faculty of Medical and Health SciencesThe University of AucklandAuckland 1023New Zealand Department of Epidemiology and Biostatistics School of Population HealthFaculty of Medical and Health SciencesThe University of AucklandAuckland 1023New Zealand

BACKGROUND Keratoconus is a degenerative corneal disease characterised by aberrant cell behaviour and loss of matrix that can result in vision *** extracted from peripheral corneas can form stem cell-enriched spheres,which have shown the potential to repopulate the normal peripheral corneal stroma in vitro upon sphere implantation but have not been previously studied in keratoconic *** To investigate the therapeutic potential of stem cell-enriched spheres formed from extracted peripheral human corneal cells when introduced to keratoconic *** Stem cell-enriched spheres were formed from extracts of normal cadaveric human peripheral corneal *** spheres were implanted into incisions created in full thickness and onto the surface of 10μm thin sections of keratoconic and normal stromal tissues in *** sections were used to maximise use of limited keratoconic tissue available for *** cells were stained with Calcein-AM and visualised with stereo and fluorescence microscopy to assess survival and behaviours between the time of implantation day 0 and 14 d(D14)from *** cells in implanted tissues were characterised for stem cell and differentiation markers using immunohistochemistry and droplet digital PCR to assess the potential implications of these characteristics in the use of spheres in keratoconus *** Spheres were successfully implanted into full-thickness central corneal tissue and onto the surface of 10μm thin en face tissue *** observable differences were seen in sphere migration,proliferation or differentiation in keratoconic tissue compared to normal between day 0 and *** stained positively with Calcein-AM up to *** migration increased from day 0 to D14,occurring radially in three dimensions from the sphere and in alignment with tissue *** proliferation marker,EdU,was detected at day *** spheres stained positively for putative stem cell markersΔNp63

关键词： Keratoconus Cell culture Immunohistochemistry Quantitative PCR Digital PCR Spheroid Holoclone Neurosphere Regeneration

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Limbal stem cells: Central concepts of corneal epithelial homeostasis

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World Journal of Stem Cells 2014年第4期6卷 391-403页

作者： Jinny J Yoon Salim Ismail trevor sherwin Department of Ophthalmology Faculty of Medical and Health SciencesUniversity of Auckland

A strong cohort of evidence exists that supports the localisation of corneal stem cells at the limbus. The distinguishing characteristics of limbal cells as stem cells include slow cycling properties, high proliferative potential when required, clonogenicity, absence of differentiation marker expression coupled with positive expression of progenitor markers, multipotency, centripetal migration, requirement for a distinct niche environment and the ability of transplanted limbal cells to regenerate the entire corneal epithelium. The existence of limbal stem cells supports the prevailing theory of corneal homeostasis, known as the XYZ hypothesis where X represents proliferation and stratification of limbal basal cells, Y centripetal migration of basal cells and Z desquamation of superficial cells. To maintain the mass of cornea, the sum of X and Y must equal Z and very elegant cell tracking experiments provide strong evidence in support of this theory. However, several recent stud-ies have suggested the existence of oligopotent stem cells capable of corneal maintenance outside of the limbus. This review presents a summary of data which led to the current concepts of corneal epithelial homeostasis and discusses areas of controversy surrounding the existence of a secondary stem cell reservoir on the corneal

关键词： Limbal stem cell Corneal epithelium XYZ hypothesis Corneal homeostasis Corneal wound repair

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