FIGURE 1.

Schematic representation of the experimental design. A, In vivo assay. Upper timeline represents the LSCD generation. Corneas were damaged with n‐heptanol followed by mechanical debridement and surgical destruction of the limbal ring. Rats were examined on days (d) 3 and 7. The procedure was repeated at d7. Transplantation was performed at d14 from the first damage. At d44, corneas were analysed by qPCR for inflammatory markers and by histology. Bottom timeline represents the culture of AT‐MSC under different conditions (standard medium, SHEM medium and CnT30 medium). AT‐MSC seeding on amniotic membrane (AM) was performed at d7. Corneal and limbal markers were assayed by qPCR. B, In vitro assay. Upper timeline shows the culture of AT‐MSC with the different media. Inflammatory markers were analysed by multiplex assay at d1, 3, 5 and 7, and by qPCR at d7. At d7 of culture, media were changed by conditioned medium (CM) of alkali‐treated HCE. AT‐MSC were cultured for 72 hours (h) with this medium. Multiplex assay was performed after 48 and 72 h from the medium change. Bottom timeline exemplifies the in vitro inflammation model by alkali treatment. NaOH treatments were applied after 24h from HCE seeding. A second round of NaOH treatment was applied after 48h from the seeding. Media were recovered at 48 and 72 h and analysed by multiplex assay for inflammatory markers. CM of alkali‐treated HCE was recovered at 72 h to culture AT‐MSC cells that were grown previously in the 3 different culture media