Pierce Jan, Benedetti Eric, Preslar Amber, Jacobson Pam, Jin Ping, Stroncek David F, Reems Jo-Anna
University of Utah Cell Therapy and Regenerative Medicine Facility, Salt Lake City, Utah.
Cell Processing Section, Department of Transfusion Medicine, Clinical Center, National Institutes of Health, Bethesda, Maryland.
Transfusion. 2017 Dec;57(12):2858-2869. doi: 10.1111/trf.14324. Epub 2017 Oct 8.
Efforts are underway to eliminate fetal bovine serum from mammalian cell cultures for clinical use. An emerging, viable replacement option for fetal bovine serum is human platelet lysate (PL) as either a plasma-based or serum-based product.
Nine industrial-scale, serum-based PL manufacturing runs (i.e., lots) were performed, consisting of an average ± standard deviation volume of 24.6 ± 2.2 liters of pooled, platelet-rich plasma units that were obtained from apheresis donors. Manufactured lots were compared by evaluating various biochemical and functional test results. Comprehensive cytokine profiles of PL lots and product stability tests were performed. Global gene expression profiles of mesenchymal stromal cells (MSCs) cultured with plasma-based or serum-based PL were compared to MSCs cultured with fetal bovine serum.
Electrolyte and protein levels were relatively consistent among all serum-based PL lots, with only slight variations in glucose and calcium levels. All nine lots were as good as or better than fetal bovine serum in expanding MSCs. Serum-based PL stored at -80°C remained stable over 2 years. Quantitative cytokine arrays showed similarities as well as dissimilarities in the proteins present in serum-based PL. Greater differences in MSC gene expression profiles were attributable to the starting cell source rather than with the use of either PL or fetal bovine serum as a culture supplement.
Using a large-scale, standardized method, lot-to-lot variations were noted for industrial-scale preparations of serum-based PL products. However, all lots performed as well as or better than fetal bovine serum in supporting MSC growth. Together, these data indicate that off-the-shelf PL is a feasible substitute for fetal bovine serum in MSC cultures.
目前正在努力从用于临床的哺乳动物细胞培养物中去除胎牛血清。人血小板裂解液（PL）作为基于血浆或血清的产品，是一种新兴的、可行的胎牛血清替代物。
进行了9次工业规模的基于血清的PL生产批次（即批次），平均±标准差体积为24.6±2.2升从单采献血者获得的汇集富血小板血浆单位。通过评估各种生化和功能测试结果对生产批次进行比较。进行了PL批次的综合细胞因子谱分析和产品稳定性测试。将用基于血浆或血清的PL培养的间充质基质细胞（MSC）的全球基因表达谱与用胎牛血清培养的MSC进行比较。
所有基于血清的PL批次中的电解质和蛋白质水平相对一致，仅葡萄糖和钙水平有轻微变化。所有9个批次在扩增MSC方面与胎牛血清一样好或更好。储存在-80°C的基于血清的PL在2年内保持稳定。定量细胞因子阵列显示基于血清的PL中存在的蛋白质既有相似之处也有不同之处。MSC基因表达谱的更大差异归因于起始细胞来源，而不是使用PL或胎牛血清作为培养补充剂。
使用大规模标准化方法，注意到工业规模制备的基于血清的PL产品存在批次间差异。然而，所有批次在支持MSC生长方面与胎牛血清一样好或更好。总之，这些数据表明现成的PL是MSC培养中胎牛血清的可行替代品。
