Limbal epithelial stem cells (LESCs) are found in the corneal limbus and are important for corneal epithelium renewal. Lack of these cells leads to a limbal stem cell deficiency (LSCD). We determined the immunogenicity of cultivated tissue from keratolimbal explants of cadaveric corneas up to the third passage grown on plastics and on intact amniotic membrane (AM). Flow cytometry was used to determine the presence of two subtypes of dendritic cells (DC), which were determined by measuring the expression of the following markers: for plasmacitoid DC (pDC): CD123+, CD303+, CD11c-, and for myeloid DC (mDC): CD11c+, CD123-. We also measured the expression of MHC II molecule, the CD80 and CD86 costumulatory molecules, and CD83 activation molecule. Populaton of mature epithelial cells was determined by measuring the expression of CK12, and the population of LESC by the expression of ABCB5. Models of forecasts of average percentage by passages were made. The difference between plastics and AM was not statistically significant (p>0.05) for any measured parameter. Increasing passages reduced the percentage of pDC and mDC, but the values were not statistically significant. The average percentage of mature epithelial corneal cells fell from first to second (p=0.007) and from second to third passage (p=0.01). The average percentage of LEMC from the first to the second passage (p=0.02) also dropped. According to our results, it is best to cultivate the cells up to and including the first passage for both substrates, thus obtaining a tissue that has a sufficient amount of epithelial corneal cells, LEMC, and a reduced proportion of mDC and pDC.
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