Yamamoto Kazuhisa, Morino Tsunetaro, Kasai Yoshiyuki, Komori Manabu, Yamato Masayuki, Kojima Hiromi
Department of Otorhinolaryngology, Jikei University School of Medicine, Tokyo, Japan.
Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University, Tokyo, Japan.
Regen Ther. 2021 Apr 7;18:59-65. doi: 10.1016/j.reth.2021.03.006. eCollection 2021 Dec.
We previously reported a new cell transplantation therapy for patients with intractable otitis media using autologous nasal mucosal epithelial cell sheets, manufactured using temperature-responsive cell culture inserts. The current study aimed to verify whether the transplantable cell sheets could be manufactured for application in clinical trials, according to standard operational procedures (SOP), in a cell processing facility (CPF).
Human nasal mucosal epithelial cells from four volunteer donors were aseptically cultured and transplantable cell sheets successfully manufactured, with reproducibility, using temperature-responsive cell culture inserts in the CPF. During the manufacture of cell sheets, the CPF environment was confirmed to be aseptic by sterilization tests. Purity of the cell sheets was confirmed by histological analysis and flow cytometry. Both safety and quality of the human nasal mucosal epithelial cell sheets were validated.
The cultured and manipulated human nasal mucosal epithelial cells showed no evidence of malignant transformation in vitro. The study confirmed the safety and suitability of the manufactured human nasal mucosal epithelial cell sheets for use in clinical trials.
The results led to the establishment of a coherent system in which transplantation could be achieved smoothly.
我们之前报道了一种针对难治性中耳炎患者的新型细胞移植疗法，该疗法使用通过温度响应性细胞培养插入物制造的自体鼻黏膜上皮细胞片。当前研究旨在验证是否可以按照标准操作程序（SOP）在细胞处理设施（CPF）中制造可用于临床试验的可移植细胞片。
对来自四名志愿者捐赠者的人鼻黏膜上皮细胞进行无菌培养，并在CPF中使用温度响应性细胞培养插入物成功制造出具有可重复性的可移植细胞片。在细胞片制造过程中，通过灭菌测试确认CPF环境无菌。通过组织学分析和流式细胞术确认细胞片的纯度。对人鼻黏膜上皮细胞片的安全性和质量均进行了验证。
培养和处理后的人鼻黏膜上皮细胞在体外未显示出恶性转化的迹象。该研究证实了所制造的人鼻黏膜上皮细胞片用于临床试验的安全性和适用性。
这些结果促成了一个连贯系统的建立，在该系统中可以顺利实现移植。
