Factors Affecting the Density of Corneal Endothelial Cells Cultured from Donor Corneas.

We investigated which specific correlation exists between the endothelial cell density (ECD) of corneal endothelial cell (CEC) cultures and the features of the donor corneas from which they originate. CEC cultures were prepared from one donor cornea or by pooling together cells of more corneas from elderly donors with ECDs lower or higher than 2000 cells/mm2. The ECDs of such primary cultures were evaluated and showed that that ECDs > 2000 cells/mm2 can be obtained only when CECs are isolated from (1) corneas of young donors; (2) at least two elderly donor corneas (if ECD > 2000 cells/mm2), or three elderly donor corneas (if ECD < 2000 cells/mm2). Secondary cultures are all characterized by ECDs < 2000 cells/mm2. Our study highlights the difficulties in obtaining cultures with ECDs > 2000 cells/mm2. Even if achievable with corneas from young donors, this becomes a challenging task when corneas from elderly donors are used (i.e., the overall majority of those collected by eye banks) and particularly when corneas from elderly donors with ECD < 2000 cells/mm2 are used. Pooling more of two corneas to obtain suitable CECs could technically overcome the problem. The above issues should be tackled appropriately before moving into clinical studies.