The proteomic and particle composition of human platelet lysate for cell therapy products

Abstract

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.

Use of flow cytometry method to detect contaminations of platelet suspensions

“…However, for bacterial flow cytometric analyzes in our study, these two solutions were found to be insufficient to lyse platelets. It was aimed to apply a platelet lysis method that can be easily used in every laboratory (Fukuda et al 2020 ; Rodrigues et al 2022 ; Kandoi et al 2018 ). In our literature review, it was seen that there were a few studies using the freeze-thaw method to achieve platelet lysis.…”