School of Medicine, Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil.
Hospital de Clínicas de Porto Alegre, Porto Alegre, Brazil.
J Cell Biochem. 2022 Sep;123(9):1495-1505. doi: 10.1002/jcb.30310. Epub 2022 Jul 26.
Following health agencies warning, the use of animal origin supplements should be avoided in biological products proposed as therapy in humans. Platelet lysate and several other growth factors sources are alternatives to replace fetal calf serum, the current gold standard in clinical-grade cell culture. However, the platelet supplement's content lacks data due to different production methods. The principle behind these products relays on the lysis of platelets that release several proteins, some of which are contained in heterogeneous granules and coordinate biological functions. This study aims to analyze the composition and reproducibility of a platelet lysate produced with a standardized method, by describing several batches' protein and particle content using proteomics and dynamic light scattering. Proteomics data revealed a diversified protein content, with some related to essential cellular processes such as proliferation, morphogenesis, differentiation, biosynthesis, adhesion, and metabolism. It also detected proteins responsible for activation and binding of transforming growth factor beta, hepatocyte growth factor, and insulin-like growth factor. Total protein, biochemical, and growth factors quantitative data showed consistent and reproducible values across batches. Novel data on two major particle populations is presented, with high dispersion level at 231 ± 96 d.nm and at 30 ± 8 d.nm, possibly being an important way of protein trafficking through the cellular microenvironment. This experimental and descriptive analysis aims to support the content definition and quality criteria of a cell supplement for clinical applications.
在卫生机构发出警告后,在拟用于人类治疗的生物制品中应避免使用动物源补充剂。血小板裂解液和其他几种生长因子来源是替代胎牛血清的替代品,胎牛血清是临床级细胞培养的当前金标准。然而,由于不同的生产方法,血小板补充剂的含量缺乏数据。这些产品背后的原理依赖于血小板的裂解,血小板释放出几种蛋白质,其中一些存在于异质颗粒中,并协调生物功能。本研究旨在通过使用蛋白质组学和动态光散射分析标准化方法生产的血小板裂解液的组成和重现性,描述几个批次的蛋白质和颗粒含量。蛋白质组学数据显示出多样化的蛋白质含量,其中一些与增殖、形态发生、分化、生物合成、粘附和代谢等重要细胞过程有关。它还检测到与转化生长因子 β、肝细胞生长因子和胰岛素样生长因子的激活和结合有关的蛋白质。总蛋白、生化和生长因子定量数据显示批次间具有一致且可重现的值。提出了两种主要颗粒群的新数据,其分散水平高达 231±96nm 和 30±8nm,可能是蛋白质通过细胞微环境进行运输的重要方式。本实验和描述性分析旨在支持临床应用细胞补充剂的内容定义和质量标准。