Jiang Yan, Fu Wei-Cai, Zhang Lin
Department of Ophthalmology, Renji Hospital Affiliated to Medical College of Shanghai Jiaotong University, Shanghai 200127, China.
Department of Ophthalmology, Renji Hospital Affiliated to Medical College of Shanghai Jiaotong University, Shanghai 200127, China.
Asian Pac J Trop Med. 2014 Aug;7(8):655-658. doi: 10.1016/S1995-7645(14)60110-3.
To explore mechanism of nduction of fibroblast to corneal endothelial cell.
Rabbit conjunctiva fibroblasts were used as feeder cells, rabbit oral mucosa epithelial cells were used as seed cells, and human denuded amniotic membrane was used as carrier to establish tissue engineering corneal endothelium. The transformation effect was observed.
As concentration of mitomycin C increased, cell survival rate gradually decreased, cell proliferation was obviously inhibited when concentration≥25 μg/mL; 5 days after being treated by 5 μg/mL mitomycin C, cell body was enlarged and extended without cell fusion, however after being treated by 0.5 μg/mL mitomycin C, cell body was significantly proliferated and gradually fused; after 3 weeks of culture, stratified epithelium appeared on rabbit oral mucosa epithelial cells, differentiation layers were 4-5 and were well differentiated, the morphology was similar to corneal endothelial cells; Under electron microscope, surface layer of cells were polygonal, tightly connected to another with microvilli on the border, there was hemidesmosome between basal cells and human denuded amniotic membrane.
Fibroblast cells have the potential of multi-directional differentiation, effective induction can promote emergence of intercellular desmosomes between seed cells and emergence of epithelial surface microvilli, and differentiate to the corneal endothelial cell. However, clinical application still needs more research and safety evaluation.
探讨诱导成纤维细胞向角膜内皮细胞转化的机制。
以兔结膜成纤维细胞为饲养层细胞，兔口腔黏膜上皮细胞为种子细胞，人脱细胞羊膜为载体构建组织工程角膜内皮，观察其转化效果。
随着丝裂霉素C浓度增加，细胞存活率逐渐降低，浓度≥25μg/mL时细胞增殖明显受抑制；5μg/mL丝裂霉素C处理5天后，细胞体积增大、伸展，无细胞融合，而0.5μg/mL丝裂霉素C处理后，细胞明显增殖并逐渐融合；培养3周后，兔口腔黏膜上皮细胞出现分层上皮，分化层数为4～5层，分化良好，形态与角膜内皮细胞相似；电镜下，细胞表层呈多边形，边界有微绒毛，细胞间紧密连接，基底细胞与脱细胞羊膜间有半桥粒。
成纤维细胞具有多向分化潜能，有效诱导可促进种子细胞间桥粒的出现及上皮表面微绒毛的出现，进而分化为角膜内皮细胞。然而，临床应用仍需更多研究及安全性评估。
