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M Chantal Hillarby, Josh Smith, Susan Shawcross, Julian Yates, Arun Brahma, Fiona Carley, Abid Ali; Can corneal limbal proteins drive the trans-differentiation of dental pulp stem cells into corneal epithelium like cells?. Invest. Ophthalmol. Vis. Sci. 2017;58(8):965.
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© ARVO (1962-2015); The Authors (2016-present)
Limbal stem cell deficiency results in the loss of normal corneal epithelial cell homeostasis and results in loss of vision. Stem cell transplantation may restore the limbal stem cells and replenish the corneal epithelium. The aim of this study was to assess whether proteins secreted by corneal limbal explants, keratinocyte growth factor (KGF) and type IV collagen (Col4), could drive Dental Pulp Stem Cells (DPSC) trans-differentiation into corneal epithelium-like cells (CEC) or precursor cells.
CD90 positive DPSC were extracted from adult teeth and cultured with 1) a non-contact co-culture with limbal explants; 2) KGF supplemented media and 3) Col4 supplemented α-MEM growth media. After eight days, inverted light microscopes was used to review histological changes that occurred in each group and immunocytochemistry and RT-PCR were conducted to detect changes in DPSC expression of mesenchymal stem cell marker CD90, CEC markers cytokeratin 12 (KRT 12) and cytokeratin 3 (KRT 3) and conjunctival markers cytokeratin 13 (CK13) and cytokeratin 19 (CK19).
DPSCs co-cultured with limbal explants or in media supplemented with either Col4 or KGF all demonstrated corneal like morphology and expression of CEC markers cytokeratin 3 and 12, consistent with the trans-differentiation into corneal epithelial like cells.
These findings may contribute to establishing a standardised culture protocol for DPSC-CEC-like trans-differentiation without relying on non-contact co-culture with limbal explants and facilitate research in to stem cell therapy for LSCD.
This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.
Immunohistochemistry staining showing cytokeratin 3 (A) and 12 (B) expression in DPSC cultured with Col4
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