Fig. 5

Effect of platelet lysates on primary neurons synaptic proteins expression and survival. Evaluation of platelet lysates on neuronal maturation was done as following (a-e). Mouse primary neurons were seeded in 12 well-plate, then treated with 0.5% (v/v) of the different platelet lysates (I-PPL, I-HPPL, HPPL) at DIV1, 3, 6, 9, 12. Whole cell lysates were prepared at DIV14 to perform Western blots to detect synaptic proteins (GluA2/3/4, Munc-18, Synatophysin or Syp and SNAP25). a Representative Western blot. (B-E) Densitometric analysis with synaptic protein levels were normalized to loading controls (β-actin). Data are given as averages from 4 experiments as percentage of the untreated controls. Evaluation of platelet lysate toxicity on mature neurons was performed as following. In addition, to evaluate impact of platelet lysates on mature neuron viability, at DIV21, cells were treated with 0.5% of the different platelet lysates and incubated for additional 2 days. The LDH level released, taken as a cytotoxic index, was then measured to determine the impact of the treatment on the viability of cells. f Schematic drawing of cells isolation method and treatment timeline. g Percentage of cytotoxicity of treated cells versus untreated controls. All data were expressed as the mean ± SD. ns: not statistically significant; **p < 0.01 vs. untreated controls. One-way analysis of variance (ANOVA) followed by Fisher’s Least Significant Difference (LSD) test