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.
Subject(s)
Biological Products , Mesenchymal Stem Cells , Somatomedins , Animals , Blood Platelets/metabolism , Cell Differentiation , Cell Proliferation , Cell- and Tissue-Based Therapy , Cells, Cultured , Culture Media/chemistry , Hepatocyte Growth Factor/metabolism , Humans , Mesenchymal Stem Cells/metabolism , Proteomics , Reproducibility of Results , Serum Albumin, Bovine/analysis , Serum Albumin, Bovine/metabolism , Somatomedins/analysis , Somatomedins/metabolism , Transforming Growth Factor beta/metabolismABSTRACT
As células-tronco mesenquimais (CTM) são consideradas células multipotentes não hematopoéticas com propriedades de autorrenovação e capacidade de diferenciação em tecidos mesenquimais e, possivelmente, em não mesenquimais. Vários estudos recentes têm reforçado o caráter multipotente destas células pela capacidade de diferenciarem-se em células derivadas da mesoderma embrionário: osteócitos, condroblastos e adipócitos. Devido ao fácil isolamento e cultivo, potencial de diferenciação e produção de fatores de crescimento e citocinas, as CTMs têm se tornado as candidatas ideais para os protocolos da medicina regenerativa. Este artigo revisa as principais características dessa célula, forma de obtenção e cultivo, propriedades imunológicas e aplicações clínicas.
Mesenchymal stem cells (MSC) are nonhematopoietic multipotent cells with selfrenewal properties and the ability to differentiate into mesenchymal tissues and possibly nonmesenchymal cells. Several lines of evidence in the past few years have confirmed the ability of these cells to differentiate into cells derived from embryonic mesoderm, such as osteocytes, adipocytes and chondroblasts. Because they are easy to isolate and culture and due to their differentiation potential and production of growth factors and cytokines, MSC have become ideal candidates for regenerative medicine protocols. This study reviews the main characteristics of MSC, how to isolate and culture them, and their immunological properties and clinical applications.
