Unidad de Producción y Reprogramación Celular, Red Andaluza Para el Diseño y Traslación de Terapias Avanzadas, Sevilla, Spain.
Unidad de Expansión y Terapia Celular. Centro de Transfusión, Tejidos y Células, Málaga, Spain.
Cytotherapy. 2021 Aug;23(8):740-753. doi: 10.1016/j.jcyt.2020.12.004. Epub 2021 Mar 11.
Successful cell cryopreservation and banking remain a major challenge for the manufacture of cell therapy products, particularly in relation to providing a hermetic, sterile cryovial that ensures optimal viability and stability post-thaw while minimizing exposure to toxic cryoprotective agents, typically dimethyl sulfoxide (MeSO).
In the present study, the authors evaluated the effectiveness and functionality of Limbo technology (Cellulis S.L., Santoña, Spain). This system provides a hermetic vial with two compartments (one for adding cells with the cryoprotective agent solution and the other for the diluent solution) and an automated defrosting device. Limbo technology (Cellulis S.L.) allows reduction of the final amount of MeSO, sidestepping washing and dilution steps and favoring standardization. The study was performed in several Good Manufacturing Practice laboratories manufacturing diverse cell therapy products (human mesenchymal stromal cells, hematopoietic progenitor cells, leukapheresis products, fibroblasts and induced pluripotent stem cells). Laboratories compared Limbo technology (Cellulis S.L.) with their standard cryopreservation procedure, analyzing cell recovery, viability, phenotype and functionality.
Limbo technology (Cellulis S.L.) maintained the viability and functionality of most of the cell products and preserved sterility while reducing the final concentration of MeSO.
Results showed that use of Limbo technology (Cellulis S.L.) offers an overall safe alternative for cell banking and direct infusion of cryopreserved cell products into patients.
成功的细胞冷冻保存和储存仍然是细胞治疗产品制造的主要挑战，特别是在提供密封、无菌的冷冻管以确保解冻后最佳活力和稳定性的同时，尽量减少接触有毒的冷冻保护剂，通常是二甲基亚砜（MeSO）。
在本研究中，作者评估了 Limbo 技术（Cellulis S.L.，西班牙桑托尼亚）的有效性和功能。该系统提供了一个带有两个隔室的密封管（一个用于添加含有冷冻保护剂溶液的细胞，另一个用于稀释剂溶液）和一个自动化解冻装置。Limbo 技术（Cellulis S.L.）允许减少最终 MeSO 的量，避免洗涤和稀释步骤，并有利于标准化。该研究在多个符合良好生产规范的实验室中进行，这些实验室制造各种细胞治疗产品（人间充质基质细胞、造血祖细胞、白细胞分离产品、成纤维细胞和诱导多能干细胞）。实验室将 Limbo 技术（Cellulis S.L.）与他们的标准冷冻保存程序进行了比较，分析了细胞回收率、活力、表型和功能。
Limbo 技术（Cellulis S.L.）保持了大多数细胞产品的活力和功能，并保持了无菌状态，同时降低了 MeSO 的最终浓度。
结果表明，使用 Limbo 技术（Cellulis S.L.）为细胞储存和直接输注冷冻保存的细胞产品提供了一种整体安全的替代方案。
