From the Schepens Eye Research Institute (U.J., L.J.), Massachusetts Eye and Ear, Boston, Massachusetts, USA.
From the Schepens Eye Research Institute (U.J., L.J.), Massachusetts Eye and Ear, Boston, Massachusetts, USA.
Am J Ophthalmol. 2022 Jul;239:244-268. doi: 10.1016/j.ajo.2022.03.015. Epub 2022 Mar 18.
Taking into consideration prior human experience with treating limbal stem cell deficiency (LSCD) with cultivated limbal epithelial cells (CLEC) from other countries, we have set a goal to optimize and standardize the techniques of CLEC preparation (called CALEC by our group) for the clinical trial in the United States.
We performed an extensive literature review of all human trials, case series, and reports involving autologous cultivated limbal epithelial cell transplantation. Allogeneic cultivated limbal epithelial cell transplantations were reported only when combined with autologous studies. We also searched prior animal data aiding in detailing regulatory toxicology requirements.
Between 1997 and 2020, the analysis of human trials revealed 21 studies on autologous grafts, and 13 studies analyzing both autologous grafts and allogeneic grafts. Of a total of 34 studies, 6 studies used good manufacturing process (GMP) facilities, and 11 studies had no animal-derived products or murine feeder layers, whereas only 1 study had both. Overall, the treatment with autologous CLEC grafts was 68.9% successful. In total there were 6 preclinical studies using rabbits, serving as surrogate studies to assess the safety and toxicity of cultivated limbal epithelial cells for human trials. Based on prior human experience, we further optimized the manufacturing conditions with GMP-grade and serum and animal-free reagents, and developed cell characterization assays for the CALEC product release.
These data were used to develop a novel and consistent manufacturing process using only qualified and validated reagents for performing the first clinical trial on CALEC transplantation to treat LSCD in the United States.
考虑到其他国家在治疗角膜缘干细胞缺乏症 (LSCD) 方面使用培养的角膜缘上皮细胞 (CLEC) 的人类经验，我们设定了一个目标，即为美国的临床试验优化和标准化 CLEC 制备技术（我们小组称之为 CALEC）。
我们对所有涉及自体培养的角膜缘上皮细胞移植的人类试验、病例系列和报告进行了广泛的文献回顾。仅当与自体研究相结合时，才报告同种异体培养的角膜缘上皮细胞移植。我们还搜索了先前有助于详细说明监管毒理学要求的动物数据。
在 1997 年至 2020 年期间，对人类试验的分析显示，有 21 项关于自体移植物的研究，以及 13 项分析自体移植物和同种异体移植物的研究。在总共 34 项研究中，有 6 项研究使用了良好生产规范 (GMP) 设施，有 11 项研究没有使用动物源性产品或鼠饲养层，而只有 1 项研究同时具备这两种条件。总体而言，自体 CLEC 移植物的治疗成功率为 68.9%。共有 6 项使用兔子的临床前研究，作为替代研究来评估培养的角膜缘上皮细胞用于人类试验的安全性和毒性。基于先前的人类经验，我们使用 GMP 级别的试剂和血清以及无动物的试剂进一步优化了制造条件，并开发了用于 CALEC 产品放行的细胞特征分析测定法。
这些数据用于开发一种新颖且一致的制造工艺，仅使用合格和经过验证的试剂，在美国进行首例使用 CALEC 移植治疗 LSCD 的临床试验。
