Platelet isolation

Platelet-rich plasma (PRP) was obtained by collecting blood in acid citrate dextrose solution (citric acid 130 mM, Trisodic citrate 152 mM, and glucose 112 mM). Samples were then diluted in Tyrode solution (HEPES 10 mM, NaCl 137 mM, KCl 2.9 mM, NaHCO3 12 mM; pH 7.4) and centrifuged at 200g for 7 minutes with deceleration at 27°C. To isolate platelets, 0.02 U/mL apyrase (A6237), 1 μM of prostaglandin E1 (P5515), and 10 μM of indomethacin (I7378-5G) provided by Sigma Aldrich, were added to PRP before being recentrifuged at 850g for 6 minutes at 27°C and suspended in Tyrode. Platelets were counted as previously described.¹⁹