Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
SHR Neuro Krebs Kardio Lipid Stoffw Microb
Schallmoser, K; Strunk, D.
Preparation of pooled human platelet lysate (pHPL) as an efficient supplement for animal serum-free human stem cell cultures.
J Vis Exp. 2009; (32): 1523-1523
Doi: 10.3791/1523
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- Führende Autor*innen der Med Uni Graz
- Schallmoser Katharina
- Strunk Dirk
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- Abstract:
- Platelet derived growth factors have been shown to stimulate cell proliferation efficiently in vivo(1,2) and in vitro. This effect has been reported for mesenchymal stromal cells (MSCs), fibroblasts and endothelial colony-forming cells with platelets activated by thrombin(3-5) or lysed by freeze/thaw cycles(6-14) before the platelet releasate is added to the cell culture medium. The trophic effect of platelet derived growth factors has already been tested in several trials for tissue engineering and regenerative therapy.(1,15-17) Varying efficiency is considered to be at least in part due to individually divergent concentrations of growth factors(18,19) and a current lack of standardized protocols for platelet preparation.(15,16) This protocol presents a practicable procedure to generate a pool of human platelet lysate (pHPL) derived from routinely produced platelet rich plasma (PRP) of forty to fifty single blood donations. By several freeze/thaw cycles the platelet membranes are damaged and growth factors are efficiently released into the plasma. Finally, the platelet fragments are removed by centrifugation to avoid extensive aggregate formation and deplete potential antigens. The implementation of pHPL into standard culture protocols represents a promising tool for further development of cell therapeutics propagated in an animal protein-free system.
- Find related publications in this database (using NLM MeSH Indexing)
- Blood Platelets - chemistry
- Cell Culture Techniques - methods
- Cell Extracts - isolation and purification
- Culture Media -
- Freezing -
- Humans -
- Mesenchymal Stem Cells - cytology
- Platelet-Derived Growth Factor - isolation and purification
- Platelet-Rich Plasma - cytology