College of Veterinary Medicine, Chungbuk National University, Cheongju, 28644, South Korea.
College of Veterinary Medicine, Kangwon National University, Chuncheon, 24341, South Korea.
Exp Eye Res. 2020 Oct;199:108192. doi: 10.1016/j.exer.2020.108192. Epub 2020 Aug 14.
Due to their very poor proliferative capacity, the dysfunction of corneal endothelial cells can sometimes lead to incurable eye diseases that require corneal transplantation. Although many studies have been performed to reconstruct corneal endothelial cells, corneal transplantation is still considered to be the established approach. In this study, we developed bio-engineered Descemet stripping endothelial (DSE) layers, using porcine cornea and induced pluripotent stem cell (iPSC)-derived corneal endothelial cells (iCECs). First, we optimized a protocol to prepare an ultra-thin and decellularized Descemet stripping (DS) scaffold from porcine cornea. Our DS layers show over 90% transparency compared to the control. Porcine-derived cells and xenogenic antigens disappeared, whereas the collagen matrix remained in the graft. Next, corneal endothelial cell lines or iCECs were seeded on the decellularized DS graft and cultured for 7 days. The drying method reduced graft rolling and edema, and increased transparency during culture. The reseeded cells were evenly distributed over the graft, and most of the cells survived. Although future clinical studies are warranted, engineered DSE tissues using xenogenic tissues and stem cells will be useful tools for the treatment of incurable corneal diseases.
由于其增殖能力极差，角膜内皮细胞的功能障碍有时会导致无法治愈的眼病，需要进行角膜移植。尽管已经进行了许多研究来重建角膜内皮细胞，但角膜移植仍然被认为是一种既定的方法。在这项研究中，我们使用猪角膜和诱导多能干细胞（iPSC）衍生的角膜内皮细胞（iCEC）开发了生物工程化的 Descemet 剥离内皮（DSE）层。首先，我们优化了从猪角膜制备超薄且去细胞化的 Descemet 剥离（DS）支架的方案。与对照相比，我们的 DS 层的透明度超过 90%。猪源性细胞和异种抗原消失，而移植物中的胶原基质仍然存在。接下来，将角膜内皮细胞系或 iCEC 接种到去细胞化的 DS 移植物上，并培养 7 天。干燥方法减少了移植物的滚动和水肿，并在培养过程中增加了透明度。重新接种的细胞均匀分布在移植物上，并且大多数细胞存活。虽然需要进行未来的临床研究，但使用异种组织和干细胞的工程化 DSE 组织将是治疗无法治愈的角膜疾病的有用工具。
