Preparation of ultra-thin descemet's membrane graft using decellularized porcine cornea
Kyungmee Park1, Hyun Kwon1, Jeonghee An1, Seo-Young Choi1, Ka-Young Lim1.
1College of Veterinary Medicine, Chungbuk National University, Cheongju, Korea
A corneal endothelial layer which is the bottom layer of cornea perform a role to maintain a clarity of cornea. When the number of corneal endothelial cell decrease due to aging or hereditary condition, it is hard to restore because its mitosis capacity is limited. As fundamental treatment, a corneal transplantation is conducted widely. However, there are lots of transplantation waiting lists through a shortage of donor cornea.
In this study, we optimize a protocol to prepare ultra-thin descemet's membrane (DM) graft using decellularized porcine cornea. Using fresh porcine eyeball, a cornea is excised including sclera and conjunctiva. With biopsy punch, endothelium, DM including stroma are partially incised. Next, the cornea was decellularized in 0.1% SDS for 5 hours. Then, a graft is detached DM including stroma. Again, the graft is decelluarized in 0.1% SDS for 10hour at room temperature. The graft was dried in dry oven at 40 celsius overnight. The induced pluripotent stem cell derived corneal endothelial cells was reseeded. Finally, a thickness of dried acellular DM graft was 57.6±7.92um and recelluarized wet graft was 99.2±5.98um. The clarity of recellularized DM graft was 91.8±2.38%.
Although the future in-vivo trials would be warranted, tissue engineering DM graft using porcine decellularized corneas might be efficient tools to resolve refractory corneal disease.
This work was supported by the Global Research and Development Center (GRDC) Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (2017K1A4A3014959) and NRF-2018R1D1A1B07050014. This work was also supported by the research grant of the Chungbuk National University in 2017..
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