The proposed study is a continuation of our Early Translational Award project (Development candidate feasibility award), which aimed to develop a xenobiotic-free culture method to expand limbal stem cells (LSCs) for transplantation. We have accomplished this goal during the previous funding period. The goal of the proposed study in this bridging funding period is to further optimize the culture condition by achieving the following two aims: 1) To optimize the culture substrate in the xenobiotic-free and feeder-free culture system; 2)To optimize the culture media of the xenobiotic-free and feeder-free culture system.
We have completed all proposed experiments and met the milestones. Methods to improve the consistency of culture substrates have been achieved and a base culture medium has been selected from three tested base media for the expansion of LSCs. The successful optimization of our LSC culture protocol allows us to proceed with the next stage of pre-IND studies.