Abstract
The search for alternatives to the use of fetal bovine serum (FBS) in cell and tissue culture media has become a major goal in terms of the 3R principles in order to reduce or to avoid harvesting of FBS from bovine fetuses, and, in terms of Good Manufacturing Practice (GMP), to ensure safe and animal product-free conditions for biomedical tissue engineering and (adult) stem cell therapy, respectively. In the present study, we investigated the feasibility of using platelet lysates (PL) as a substitute for FBS, based on the fact that most of the potent mitogenic factors present in serum are derived from activated thrombocytes. Platelet lysates were obtained from outdated human donor platelet concentrates. Methods were established to activate human donor platelets in order to achieve a maximum yield of platelet a-granule growth factors. Platelet lysates were successfully introduced to grow and maintain anchorage-dependent and -independent human and animal cell lines. For cell culture experiments, cells were either grown in culture media supplemented with 10% FBS, 5% PL, or under serum-free conditions. Growth experiments, viability assays, and platelet lysate-induced activation of ERK1/2 mitogen-activated protein kinase revealed platelet lysates as a valuable alternative to FBS in cell culture media.
Highlights
Propagation of human and animal cells in vitro requires adequate culture conditions and culture media
Basal media are frequently supplemented with animal serum, mostly fetal bovine serum (FBS), which promotes cell growth and proliferation (Gstraunthaler, 2003; Lindl und Gstraunthaler, 2008)
Preparation of human platelet lysates Outdated human donor platelets obtained by different apheresis methods (AmicusTM, Fenwal, Lake Zurich, IL, USA; TrimaTM, CaridianBCT, Lakewood, CO, USA) were provided by the blood bank of the Innsbruck Medical University Hospital
Summary
Propagation of human and animal cells in vitro requires adequate culture conditions and culture media. The culture medium has to supply all essential nutrients for cell metabolism, growth and proliferation. Basal media are frequently supplemented with animal serum, mostly fetal bovine serum (FBS), which promotes cell growth and proliferation (Gstraunthaler, 2003; Lindl und Gstraunthaler, 2008). The major functions of serum in culture media are to provide hormonal factors stimulating cell growth and proliferation, to promote differentiated functions, and to provide transport proteins, minerals, trace elements, lipids, attachment and spreading factors, and stabilizing and detoxifying factors needed for maintaining pH or to inhibit proteases either directly, such as α-antitrypsin, or indirectly, by acting as an unspecific sink for proteases and other (toxic) molecules. FBS is superior to serum from adult animals because of its low gamma-globulin content, as a high content of antibodies may inhibit growth and proliferation
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