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Since 1997, LSCD has been treated successfully by transplanting cultured limbal epithelial stem cells from donors.In bilateral LSCD, limbal tissue can be provided from a relative or a deceased individual, however, any non-autologous source requires prolonged immunosuppressive treatment.Furthermore, cholera toxin and epidermal growth factors, which are typically added to DMEM, increase the proliferation capacity of oral epithelial cells, we hypothesized that the harvesting site could affect the growth capacity and phenotype of ex vivo expanded OMECs.

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Since 2010, following a study by Rama et al., the degree of differentiation of cultured epithelia for treating LSCD has been a major issue in corneal regenerative medicine.

Rama and colleagues demonstrated that transplantation of undifferentiated limbal epithelial sheets yields significantly better clinical results compared to the use of more differentiated equivalents.

The submucosal connective tissue was removed by dissection using forceps, scalpel, and a dissection microscope (Leica ZOOM 200, Leica Microsystems Inc., Buffalo, IL).

The tissue samples were cut into 1–3 mm explants and immersed in the various media containing antibiotics (50 IU/ml P/S).

All experiments using the animal were carried out in accordance with the approved guidelines.