Human Platelet Lysates as a Serum Substitute in Cell Culture Media

The search for alternatives to fetal bovine serum (FBS) has become a major goal in the field of cell and tissue culture research. Although the supplementation of culture media with FBS is routine practice, FBS bears a number of disadvantages: unknown composition, high lot-to-lot variablity, ethical concerns about the harvest from bovine fetuses, and possible shortage in global supply. Several strategies have been developed to reduce or replace FBS in cell culture media (Bjare 1992; Even et al. 2006; Gstraunthaler 2003; van der Valk et al. 2004, 2010). Here we report on the use of human platelet lysates (PL) as a serum replacement (Alden et al. 2007; Bernardo et al. 2007; Bieback et al. 2009; Doucet et al. 2005; Johansson et al. 2003; Kocaoemer et al. 2007; Muller et al. 2009; Schallmoser et al. 2009). PL in DMEM support growth, proliferation and differentiation, as assessed by dome formation, of proximal tubule-like LLC-PK1 (porcine kidney) and HK-2 (human kidney) cells, as well as PL-supplemented DMEM/Ham F-12 for distal tubule -like MDCK (dog kidney) cells. In addition to adherent epithelial cell lines, anchorage-independent Raji human lymphoma cells were investigated. PL fully supported growth and proliferation of Raji cells in RPMI-1640 medium in suspension. In order to biochemically deter nine the proliferative potential of PL, the stimulation of extracellular signal-regulated MAP kinase (ERK1/2) was determined. Addition of PL to quiescent LLC-PK1 cultures resulted in specific phosphorylation, and thus activation, of ERK1/2 within minutes. The time course is identical with ERK1/2 activation upon addition of FBS. The data show the high potential of PL as a valuable substitute for FBS in mammalian cell and tissue culture.