根据 GMP 级方案扩增间充质基质细胞中的人血小板裂解物：细胞工厂的经验。
Human platelet lysate in mesenchymal stromal cell expansion according to a GMP grade protocol: a cell factory experience.
机构信息
Cell Factory Meyer, "A. Meyer" University Children's Hospital, Florence, Italy.
Transfusion Medicine and Cell Therapy Unit, "A. Meyer" University Children's Hospital, Florence, Italy.
出版信息
Stem Cell Res Ther. 2018 May 2;9(1):124. doi: 10.1186/s13287-018-0863-8.
BACKGROUND
The use of platelet lysate (PL) for the ex-vivo expansion of mesenchymal stromal/stem cells (MSCs) was initially proposed by Doucet et al. in 2005, as an alternative to animal serum. Moreover, regulatory authorities discourage the use of fetal bovine serum (FBS) or other animal derivatives, to avoid risk of zoonoses and xenogeneic immune reactions. Even if many studies investigated PL composition, there still are some open issues related to its use in ex-vivo MSC expansion, especially according to good manufacturing practice (GMP) grade protocols.
METHODS
As an authorized cell factory, we report our experience using standardized PL produced by Azienda Ospedaliero Universitaria Meyer Transfusion Service for MSC expansion according to a GMP grade clinical protocol. As suggested by other authors, we performed an in-vitro test on MSCs versus MSCs cultured with FBS that still represents the best way to test PL batches. We compared 12 MSC batches cultured with DMEM 5% PL with similar batches cultured with DMEM 10% FBS, focusing on the MSC proliferation rate, MSC surface marker expression, MSC immunomodulatory and differentiation potential, and finally MSC relative telomere length.
RESULTS
Results confirmed the literature data as PL increases cell proliferation without affecting the MSC immunophenotype, immunomodulatory potential, differentiation potential and relative telomere length.
CONCLUSIONS
PL can be considered a safe alternative to FBS for ex-vivo expansion of MSC according to a GMP grade protocol. Our experience confirms the literature data: a large number of MSCs for clinical applications can be obtained by expansion with PL, without affecting the MSC main features. Our experience underlines the benefits of a close collaboration between the PL producers (transfusion service) and the end users (cell factory) in a synergy of skills and experiences that can lead to standardized PL production.
背景
血小板裂解液（PL）最初由 Doucet 等人于 2005 年提出，作为动物血清的替代品，用于间充质基质/干细胞（MSCs）的体外扩增。此外，监管机构不鼓励使用胎牛血清（FBS）或其他动物衍生产品，以避免人畜共患病和异种免疫反应的风险。即使许多研究调查了 PL 的组成，但在根据良好生产规范（GMP）级别的协议进行体外 MSC 扩增方面，仍然存在一些悬而未决的问题。
方法
作为授权的细胞工厂，我们报告了使用 Azienda Ospedaliero Universitaria Meyer 输血服务部生产的标准化 PL 根据 GMP 级别的临床方案进行 MSC 扩增的经验。正如其他作者所建议的，我们对 MSCs 与用 FBS 培养的 MSCs 进行了体外测试，这仍然是测试 PL 批次的最佳方法。我们比较了用 DMEM 5% PL 培养的 12 个 MSC 批次与用 DMEM 10% FBS 培养的类似批次，重点关注 MSC 的增殖率、MSC 表面标志物表达、MSC 免疫调节和分化潜能，以及最终的 MSC 相对端粒长度。
结果
结果证实了文献数据，PL 增加了细胞增殖，而不影响 MSC 的免疫表型、免疫调节潜能、分化潜能和相对端粒长度。
结论
根据 GMP 级别的方案，PL 可被视为 FBS 用于 MSC 体外扩增的安全替代品。我们的经验证实了文献数据：通过 PL 扩增，可以获得大量用于临床应用的 MSC，而不影响 MSC 的主要特征。我们的经验强调了 PL 生产商（输血服务）和最终用户（细胞工厂）之间密切合作的好处，这种合作可以实现技能和经验的协同，从而导致 PL 的标准化生产。
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