About the project:

The project’s main goal is to introduce the treatment for limbal stem cell deficiency (LSCD) into clinical practice. Under experimental conditions, we culture (grow and multiply) limbal epithelial cells obtained from the eye, which will be used to treat unilateral LSCD, and epithelial cells from the oral mucosa, used to treat bilateral LSCD. We prepare and test suitable materials (polylactide nanofibers, fibrin glue) on which the cells grow and the solutions in which we culture them. Additionally, we are developing tests to evaluate the DNA stability of cultured cells and their long-term storage.


In 2021, we focused on preparing limbal stem cells from small segments of limbal tissue obtained from the surface of the eye. These correspond to the size of tissue that can be obtained from patients. We started culturing the cells obtained from the oral mucosa on different carriers and in different culture media. Unlike limbal stem cells, for which we prepare explant cultures, it seems more appropriate to use a cell suspension for culturing cells from the buccal mucosa. The situation associated with the Covid 19 pandemic caused many problems (low number of oral mucosal tissue samples and absence of clean rooms for the project). We addressed the situation by focusing experimental work on available limbal cells (obtained from excess tissue after corneal transplantation). We experimentally found that their proliferation, necessary for the survival of the graft after transplantation, is strongly supported by interleukin 13. We are also looking for and testing the most suitable markers to verify the stemness of the cells, which is a prerequisite for successful clinical results.In addition, we are developing assays for to assess the stability of cultured cell DNA and its long-term storage. In conclusion, we can summarize that, despite the Covid pandemic, the project is running and bringing the first results.


We have developed and standardized a method of culturing limbal epithelial cells (cells obtained from the area between the cornea and conjunctiva, where the stem cells for the corneal epithelium are located) using an explant culture. This means the cells grow and proliferate, expanding from a small piece of limbal tissue under suitable conditions (fibrin glue was used as a feeder layer for cell growth in solution with additives ensuring cell nutrition). We are working on finding the most suitable conditions for culturing cells from the buccal mucosa. A cell suspension cultured on fibrin glue appears to be the most suitable (with the highest number of stem cells). As an even more effective alternative, we are testing an amniotic membrane, which could increase the stemness of the prepared cells, and polylactide nanofibers coated with fibronectin, which supports the attachment of cultured cells. Last year, we acquired clean rooms for the project. Negotiations on their necessary modification and the transfer of cultivations from experimental to tissue bank conditions are ongoing with the State Institute for Drug Control.

Beneficiary and project partners:

Univerzita Karlova
University of Oslo
NILU-Norwegian Institute for Air Research
Ústav makromolekulární chemie AV ČR, v. v. i.
Fakultní nemocnice Královské Vinohrady

The EYEFORTX_2 project benefits from a € 1 mil. grant from Norway Grants and Technology Agency of the Czech Republic. The project is carried out under the KAPPA funding programme for applied research, experimental development and innovation, managed by the Technology Agency of the Czech Republic.