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首页> 外文期刊>Frontiers in Cell and Developmental Biology >A Blood Bank Standardized Production of Human Platelet Lysate for Mesenchymal Stromal Cell Expansion: Proteomic Characterization and Biological Effects
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A Blood Bank Standardized Production of Human Platelet Lysate for Mesenchymal Stromal Cell Expansion: Proteomic Characterization and Biological Effects

机译：用于间充质基质细胞扩张的人血小板裂解物的血库标准化生产：蛋白质组学特征和生物学作用

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Human Platelet Lysate (hPL) is considered a valid substitute to foetal bovine serum (FBS) in mesenchymal stromal cells (MSC) expansion and it is commonly produced starting from intermediate side-products of whole blood donations. Through freeze-thaw cycles, hPL is highly enriched in chemokines, growth factors and adhesion and immunologic molecules. Cell therapy protocols, using hPL instead of FBS for cells' expansion, are approved by regulatory authorities without concerns and administration in patients is considered safe. However, published data are fairly difficult to compare, since the production of hPL is highly variable. This study proposes to optimize and standardize the hPL productive process by using instruments, technologies and quality/safety standards required for Blood Banks activities and products. The quality and improved selection of the starting material (i.e. the whole blood), together with the improvement of the production process, guarantee a product characterized by higher content and quality of growth factors as well as a reduction in batches' variability. By increasing the number of freeze/thaw cycles from one (hPL1c) to four (hPL4c), we obtained a favourable effect on the release of growth factors from platelet ? granules. Those changes have directly translated into biological effects leading to a decreasing Doubling Time (DT) of MSC expansion at 7 days (49.41±2.62 hours vs 40.61±1.11 hours, P0.001). Furthermore, Mass Spectrometry (MS)-based evaluation has shown that the proliferative effects of hPL4c are also combined with a lower batch-to-batch variability (10-15% vs 21-31%) at the proteomic level. In conclusion, we have considered lot-to-lot hPL variability and, by the strict application of Blood Bank standards, we have obtained a standardized, reproducible, safe, cheap and ready-to-use product.
机译：人的血小板裂解物（HPL）被认为是胎儿基质细胞（MSC）膨胀中的胎牛血清（FBS）的有效替代物，并且通常从整个献血的中间副产品开始。通过冻融循环，HPL高度富含趋化因子，生长因子和粘附和免疫分子。使用HPL代替FBS的细胞疗法方案用于细胞的扩张，经监管当局批准，不担忧，患者的施用被认为是安全的。然而，由于HPL的生产是高度变量，所公布的数据相当难以比较。本研究建议通过使用血库活动和产品所需的仪器，技术和质量/安全标准来优化和标准化HPL生产过程。起始材料的质量和改进的选择（即全血）以及改进生产过程，保证了一种产品，其特征在于生长因子的更高内容和质量，以及批量变异性的减少。通过将来自一种（HPL1C）至四（HPL4C）的冻结/解冻循环循环的数量增加，我们对血小板的生长因子释放获得了有利影响？颗粒。这些变化直接翻译成生物效应，导致MSC扩增的倍增时间（DT）在7天内（49.41±2.62小时Vs 40.61±1.11小时，P <0.001）。此外，基于质谱（MS）的评价表明，HPL4C的增殖效应也与蛋白质组学水平的较低的间歇到分批变异性（10-15％vs 21-31％）结合。总之，我们已经考虑了批量批次的HPL变异性，并且由于严格应用血库标准，我们已获得标准化，可重复，安全，便宜和即用的产品。

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