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
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