大鼠脑微血管内皮细胞的原代培养 - 第三军医大学学报.DOC

角膜缘上皮细胞与冻干的脱细胞猪角膜基质修复碱烧伤兔角膜的实验研究 郭梁，刘恺鸣，胡丹 (第四军医大学西京医院眼科，全军眼科研究所，西安 710032) 摘要：目的 探讨应用角膜缘上皮细胞(limbal epithelial cells，LECs)冻干脱细胞猪角膜基质(freeze-dried decellularized porcine corneal stroma, freeze-dried DPCS) 修复碱烧伤。方法 培养制备冻干的将两者复合培养后板层移植到碱烧伤的兔角膜，观察移植后2月内的角膜修复情况。结果 免疫细胞化学RT-PCR检测培养的中有表达ABCG2的LSCs。大体观察移植后角膜混浊度减轻，新生血管消退。HEMasson Trichrome染色显示移植后上皮面的细胞覆层化生长炎性细胞浸润减少DPCS纤维整合于碱烧伤的角膜植床，角膜胶原排列趋于透射电镜检查。结论 联合冻干的移植能够修复碱烧伤的兔眼角膜。 ：角膜缘上皮细胞；冻干的脱细胞猪角膜基质；碱烧伤 R318.08; R779.65; Q813.11 文献标识码：A Management of corneal alkali burn with limbal epithelial cells and freeze-dried decellularized porcine corneal stroma transplantation in a rabbit model GUO Liang, LIU Kaiming, HU Dan (Department of Ophthalmology, Xijing Hospital, Fourth Military Medical University, Xi’an 710032, China) Abstract: Objective To investigate the therapeutic effect of limbal epithelial cells-freeze-dried decellularized porcine corneal stroma construct transplantation in rabbits with corneal alkali burn. Methods Allogeneic rabbit limbal epithelial cells were cultured and identified. Porcine cornea scaffold was prepared by decellularized and freeze-dried process.The LECs-DPCS construct was cultured and transplanted to the alkali burned corneal surface in rabbits for 2 months follow-up. Results Immunocytochemistry and RT-PCR identification revealed the presence of ABCG2 positive limbal stem cells. General view showed the postoperative corneas were clear with few new vessels. HE and Masson Trichrome staining results indicated the epithelium stratification, fewer cellular infiltration, DPCS’ integration into host corneal tissues, and more regular collagen configuration. Transmission electron microscopy exhibited formation of desmosome and microvilli. Conclusion Allogeneic limbal epithelial cells transplantation with freeze-dried DPCS as a carrier can be used to restore corneal clarity after alkali burn in a rabbit model. Key words: limbal epithelial cells; freeze-dried decellularized porci