Sphere-forming cells from peripheral cornea demonstrate the ability to repopulate the ocular surface
Autor: | Trevor Sherwin, C. N. J. McGhee, Salim Ismail, Jeremy J Mathan, Jennifer J. McGhee |
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Rok vydání: | 2016 |
Předmět: |
0301 basic medicine
Pathology genetic structures Cellular differentiation Medicine (miscellaneous) Gene Expression Stem cell marker Holoclone Tissue Culture Techniques Cornea Quantitative PCR 0302 clinical medicine Cell Movement ATP Binding Cassette Transporter Subfamily G Member 2 Limbal stem cell Receptor Notch1 Stem Cells Epithelium Corneal Cell Differentiation Neoplasm Proteins medicine.anatomical_structure Molecular Medicine Proteoglycans Stem cell Immunocytochemistry medicine.medical_specialty Stromal cell ATP Binding Cassette Transporter Subfamily B Biology Limbus Corneae Biochemistry Genetics and Molecular Biology (miscellaneous) 03 medical and health sciences Proliferating Cell Nuclear Antigen Spheroids Cellular medicine Cadaver Humans Vimentin ATP Binding Cassette Transporter Subfamily B Member 1 Progenitor cell Cell Proliferation Tumor Suppressor Proteins Research Cell Biology eye diseases 030104 developmental biology Spheroid 030221 ophthalmology & optometry sense organs Keratin-3 Laminin Cell culture Keratocan Biomarkers Stem Cell Transplantation Transcription Factors |
Zdroj: | Stem Cell Research & Therapy |
ISSN: | 1757-6512 |
Popis: | Background The limbus forms the outer rim of the cornea at the corneoscleral junction and harbours a population of stem cells for corneal maintenance. Injuries to the limbus, through disease or accidents such as chemical injuries or burns, may lead to significant visual impairment due to depletion of the native stem cells of the tissue. Methods Sphere-forming cells were isolated from peripheral cornea for potential use as transplantable elements for limbal stem cell repopulation and limbal reconstruction. Immunocytochemistry, live cell imaging and quantitative PCR were used to characterize spheres and elucidate activity post implantation into human cadaveric corneal tissue. Results Spheres stained positively for stem cell markers ∆NP63α, ABCG2 and ABCB5 as well as the basal limbal marker and putative niche marker, notch 1. In addition, spheres also stained positively for markers of corneal cells, vimentin, keratin 3, keratocan and laminin, indicating a heterogeneous mix of stromal and epithelial-origin cells. Upon implantation into decellularized corneoscleral tissue, 3D, polarized and radially orientated cell migration with cell proliferation was observed. Cells migrated out from the spheres and repopulated the entire corneal surface over 14 days. Post-implantation analysis revealed qualitative evidence of stem, stromal and epithelial cell markers while quantitative PCR showed a quantitative reduction in keratocan and laminin expression indicative of an enhanced progenitor cell response. Proliferation, quantified by PCNA expression, significantly increased at 4 days subsequently followed by a decrease at day 7 post implantation. Conclusion These observations suggest great promise for the potential of peripheral corneal spheres as transplantable units for corneal repair, targeting ocular surface regeneration and stem cell repopulation. |
Databáze: | OpenAIRE |
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