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HPL Culture System – Two-phase cell culture system based upon human platelet lysate
In order to provide cells with a sufficient amount of growth factors, they are usually cultured in medium supplemented with fetal calve serum (FCS) or other blood sera. In the case of therapeutic applications, these xenogenic components bear the risk of immunologic reactions and infection with viruses or prions. Human platelet lysate (HPL) is considered to be an attractive alternative to FCS. It has been shown that HPL-supplemented cell culture medium yields a high expansion rate and is superior to FCS even during longterm cultivation.
Still, cells covered with HPL-medium do not experience optimal growth conditions, as they are usually cultured on a plastic surface which involves contact inhibition and enzymatic treatment during passaging. Invention :This invention relates to a cell culture system based upon human platelet lysate (HPL) and standard cell culture medium. The two-phase system consists of HPL-medium and HPL-gel. Both components only differ in the addition of a polymerization-inhibiting substance, e.g. heparin (compare Fig.1). In this way, growth factors are provided in a constant concentration throughout the culture system and three-dimensional cell growth is facilitated, thus minimizing contact inhibition. It has been shown that culturing mesenchymal stromal cells (MSC) on HPL-gel yields a higher proliferation rate (compare Fig. 3) and CFU-f outgrowth compared to a plastic surface and collagen-gel. Cell passaging can either be performed enzymatically or by mechanical gel fragmentation with a pipette (compare Fig. 2). Summing up, the novel HPL culture system provides excellent growth conditions for standard cell culture as well as for cell therapeutic applications.
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