Utheim Tor Paaske, Utheim Øygunn Aass, Khan Qalb-E-Saleem, Sehic Amer
Department of Medical Biochemistry, Oslo University Hospital, Oslo 0407, Norway.
Department of Oral Biology, Faculty of Dentistry, University of Oslo, Oslo 0372, Norway.
J Funct Biomater. 2016 Mar 1;7(1):5. doi: 10.3390/jfb7010005.
The cornea is critical for normal vision as it allows allowing light transmission to the retina. The corneal epithelium is renewed by limbal epithelial cells (LEC), which are located in the periphery of the cornea, the limbus. Damage or disease involving LEC may lead to various clinical presentations of limbal stem cell deficiency (LSCD). Both severe pain and blindness may result. Transplantation of cultured autologous oral mucosal epithelial cell sheet (CAOMECS) represents the first use of a cultured non-limbal autologous cell type to treat this disease. Among non-limbal cell types, CAOMECS and conjunctival epithelial cells are the only laboratory cultured cell sources that have been explored in humans. Thus far, the expression of p63 is the only predictor of clinical outcome following transplantation to correct LSCD. The optimal culture method and substrate for CAOMECS is not established. The present review focuses on cell culture methods, with particular emphasis on substrates. Most culture protocols for CAOMECS used amniotic membrane as a substrate and included the xenogeneic components fetal bovine serum and murine 3T3 fibroblasts. However, it has been demonstrated that tissue-engineered epithelial cell sheet grafts can be successfully fabricated using temperature-responsive culture surfaces and autologous serum. In the studies using different substrates for culture of CAOMECS, the quantitative expression of p63 was generally poorly reported; thus, more research is warranted with quantification of phenotypic data. Further research is required to develop a culture system for CAOMECS that mimics the natural environment of oral/limbal/corneal epithelial cells without the need for undefined foreign materials such as serum and feeder cells.
角膜对于正常视力至关重要,因为它能使光线传导至视网膜。角膜上皮由位于角膜周边即角膜缘的角膜缘上皮细胞(LEC)更新。涉及LEC的损伤或疾病可能导致角膜缘干细胞缺乏症(LSCD)的各种临床表现。严重疼痛和失明都可能由此产生。培养的自体口腔黏膜上皮细胞片(CAOMECS)移植是首次使用培养的非角膜缘自体细胞类型来治疗这种疾病。在非角膜缘细胞类型中,CAOMECS和结膜上皮细胞是仅有的在人体中探索过的实验室培养细胞来源。迄今为止,p63的表达是移植矫正LSCD后临床结果的唯一预测指标。CAOMECS的最佳培养方法和基质尚未确定。本综述聚焦于细胞培养方法,尤其着重于基质。大多数CAOMECS的培养方案使用羊膜作为基质,并包含异种成分胎牛血清和鼠3T3成纤维细胞。然而,已证明使用温度响应性培养表面和自体血清能够成功制备组织工程化上皮细胞片移植物。在使用不同基质培养CAOMECS的研究中,p63的定量表达普遍报道较少;因此,需要更多关于表型数据定量的研究。需要进一步研究来开发一种用于CAOMECS的培养系统,该系统能模拟口腔/角膜缘/角膜上皮细胞的自然环境,而无需血清和饲养细胞等不确定的外来物质。