To evaluate the effectiveness of cultured human limbal epithelium for the treatment of limbal stem cell deficiency

角膜是眼睛前部的透明窗口，它的清晰度对光传输到眼睛后部的视觉感知至关重要。角膜的透明度取决于其最外层上皮的完整性，在整个生命过程中，在角膜边缘被称为边缘的一群干细胞不断再生，即所谓的边缘干细胞（LSC）。如果LSC由于受伤或疾病而变得缺乏，这种上皮将会破裂，导致严重的疼痛和失明。这种情况通常难以治疗，然而，近年来，角膜缘干细胞缺乏症（LSCD）的治疗取得了重大突破。病人的LSC ？自己的（单眼疾病）或相对健康的眼睛或来自尸体的眼睛可以在实验室中生长，然后移植到患病的眼睛上。然而，这种技术使用动物产品来帮助细胞生长，有很大的疾病传播风险。此外，由于报告病例的多样性和数量少，这种治疗的安全性和有效性很难确定。因此，在正式的结构化临床试验确定其有效性和安全性之前，培养的角膜缘上皮移植不能被认可，从而使NHS能够资助。这种试验是本建议的主要目标。因此，我们开发了一种？不含动物产品？在实验室超净环境中培养细胞的技术，并成功地从健康的另一只眼睛中取出LSC，移植了8名LSCD患者的一只眼睛。尽管我们取得了成功，但这个病人的数量不足以得出正确的结论。因此，本建议旨在通过治疗更多的单侧LSCD患者来评估我们的方法的安全性和有效性。该项目的另一个目标是开发一种冷冻方法来保存培养的人类LSC，并建立一项服务，为英国其他中心的移植提供临床级培养的LSC。此外，本研究将调查LSCD患者的异常泪膜，以便定制眼部治疗。最后，LSCD对患者有巨大的影响。对生产力和休闲活动造成严重的不利影响。为此，我们选择了4份问卷来研究LSCD患者及其治疗的不同方面。

The cornea is the clear window at the front of the eye and its clarity is vital for light transmission to the back of the eye for visual perception. Transparency of the cornea depends upon the integrity of its outermost layer called the epithelium, which is constantly regenerated throughout life by a population of stem cells found at the edge of the cornea in an area called the limbus, the so called limbal stem cells (LSC). If LSC becomes deficient as a result of injury or disease, this epithelium will breakdown leading to severe pain and blindness. This condition is often difficult to treat, however, the management of limbal stem cell deficiency (LSCD) has benefited from major breakthroughs in recent years. LSC from the patient?s own (one eye disease) or a relative healthy eye or from a cadaveric eye can grow in the laboratory and then transplanted to the diseased eye. This technique however uses animal products to help grow the cells with a great risk of diseases transmission. In addition, the safety and effectiveness of this treatment is rather difficult to establish due to the diversity and the small number of cases reported. Transplantation of cultured limbal epithelium can therefore not be recognized, enabling NHS funding, until its effectiveness and safety is determined by a formally structured clinical trial. Such a trial forms the main objective of this proposal. We have therefore developed an ?animal product free? technique for growing the cells in the laboratory in an ultra-clean environment and successfully transplanted 8 patients with LSCD in one eye by taking LSC from their healthy other eye. Despite of our success, this number of patients is not significant to draw proper conclusions. This proposal aims therefore to evaluate the safety and efficacy of our methods by treating more patients with total unilateral LSCD. Another aim of the project is to develop a freezing method of cultured human LSC for storage and to set-up a service to supply clinical grade cultured LSC for transplantation at other UK centres. In addition, this study will investigate the abnormal tear film in patients with LSCD to allow for a customised eye treatment. Finally, LSCD has an enormous impact on patient?s quality of life with serious detrimental effects on productivity and leisure activities. To that effect we have selected 4 questionnaires to look at different aspects of patients with LSCD and its treatment.