Reproducibility of GMP-compliant production of therapeutic stressed peripheral blood mononuclear cell-derived secretomes, a novel class of biological medicinal products
Reproducibility of GMP-compliant production of therapeutic stressed peripheral blood mononuclear cell-derived secretomes, a novel class of biological medicinal products
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DOI：
10.1186/s13287-019-1524-2
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发表时间：
2020-01-03
影响因子：
7.5
通讯作者：
Ankersmit, Hendrik Jan
中科院分区：
文献类型：
--
作者：
Laggner, Maria;Gugerell, Alfred;Ankersmit, Hendrik Jan
背景最近的概念，基于分泌组的组织再生已经深刻地改变了再生医学领域，并提供了有前途的新的治疗选择。与具有单一活性物质的药品相比，细胞源性分泌体包含多效性生物活性成分，这是可重现的药品有效性和患者安全性的主要障碍。符合药品生产质量管理规范（GMP）的生产保证了生物药品的高批间一致性和可重现的疗效，但GMP下生产的不同批次细胞分泌体尚未进行比较，也未建立合适的质量控制参数。为此，我们分析了在GMP下生产的不同批次的分泌组的各种生物学和功能参数，所述分泌组从经γ-辐照的外周血单核细胞获得，所述外周血单核细胞在梗塞心肌、中风、脊髓损伤和皮肤伤口中具有经证实的组织再生特性。离体主动脉环发芽测定和基于细胞的蛋白质和报道基因测定。此外，我们确定分泌体的稳定性在不同的批次后6个月的存储在不同的环境temperature.ResultsWe观察到，批间差异的生物活性成分和分泌体的属性是小的，尽管相当大的差异，蛋白质浓度和效力之间的个别捐助者分泌体。稳定性测试表明，当冻干物在+5 ℃下储存6个月时，分泌体的分析和功能性质保持稳定。结论我们首次证明了细胞来源的无细胞分泌体作为生物医药产品的一致性生产。本研究的结果为符合GMP的治疗性细胞分泌组生产选择适当的质量控制参数提供了基础，并为未来在组织再生医学中使用分泌组的临床试验铺平了道路。
BackgroundThe recent concept of secretome-based tissue regeneration has profoundly altered the field of regenerative medicine and offers promising novel therapeutic options. In contrast to medicinal products with a single active substance, cell-derived secretomes comprise pleiotropic bioactive ingredients, representing a major obstacle for reproducible drug product efficacy and warranting patient safety. Good manufacturing practice (GMP)-compliant production guarantees high batch-to-batch consistency and reproducible efficacy of biological medicinal products, but different batches of cellular secretomes produced under GMP have not been compared yet, and suitable quality control parameters have not been established. To this end, we analyzed diverse biological and functional parameters of different batches produced under GMP of the secretome obtained from gamma -irradiated peripheral blood mononuclear cells with proven tissue regenerative properties in infarcted myocardium, stroke, spinal cord injury, and skin wounds.MethodsWe quantified key secretome ingredients, including cytokines, lipids, and extracellular vesicles, and functionally assessed potency in tube formation assay, ex vivo aortic ring sprouting assay, and cell-based protein and reporter gene assays. Furthermore, we determined secretome stability in different batches after 6months of storage at various ambient temperatures.ResultsWe observed that inter-batch differences in the bioactive components and secretome properties were small despite considerable differences in protein concentrations and potencies between individual donor secretomes. Stability tests showed that the analytical and functional properties of the secretomes remained stable when lyophilisates were stored at temperatures up to +5 degrees C for 6months.ConclusionsWe are the first to demonstrate the consistent production of cell-derived, yet cell-free secretome as a biological medicinal product. The results from this study provide the basis for selecting appropriate quality control parameters for GMP-compliant production of therapeutic cell secretomes and pave the way for future clinical trials employing secretomes in tissue regenerative medicine.