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Ricardo Pedro Casaroli-Marano, Eva M Martínez-Conesa, Nuria Nieto-Nicolau, Francisco Arnalich-Montiel, Sherezade Fuentes-Julian, Maria P De Miguel; Adipose Derived Stem Cells (ADS) for Ocular Surface Regeneration.. Invest. Ophthalmol. Vis. Sci. 2014;55(13):5183.
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The use of stem cells is promising for future cell-based therapies such as tissue regeneration and engineering. Mesenchymal stem cells (MSCs) derived from adult human adipose tissue (ADS) manifest multilineage differentiation capacity. However, their plasticity to differentiate into epithelial characteristics still remains little well-known. The corneal epithelium is maintained by limbal stem cells. Transplantation of autologous corneal stem cells it is not possible in several cases in which bilateral disease produces total corneal stem-cell deficiency. For this reason, the use of autologous ADS as a source of cells for the ocular surface reconstruction is being studied.
Pool of ADS cells from different healthy donors was generated. The plasticity, cellular and molecular characterization and ADS in vitro behavior to assumes epithelial-like characteristic were studied. Several limbic cell markers (p63, ABCG2, NGFr) and a set of epithelial markers (cytokeratins 3 and 12, integrins, EGFr and E-cadherin) have been analyzed by RT-PCR methods. After five days in cultures ADS cells were seeded onto human amniotic membrane and applied in a rat model of limbic deficiency. Histopathological and molecular analyses were carried-out between 21 and 28 days.
ADS cells can assume epithelial-like phenotypes when cultured into CnT-30 or SHEM media after 5 days in culture. Expression of an epithelial marker set was more evident in SHEM between 3rd and 7th days in culture. Histopathological studies revealed restructuring of corneal surface, more evident with ADS cells cultured in SHEM. PCR experiments with sequencing methods demonstrated the presence of human CK12 in ADS cells and in corneas of murine in vivo model.
ADS cells could be a potential cell type for ocular surface regenerative medicine, and the epithelial regeneration and the expression of CK12 in murine model of limbic deficiency after 3 weeks may support this hypothesis.
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