Simple and rapid isolation of platelets using Ficoll-Uropoline cushion.

A simplified procedure for separation of human platelets from plasma proteins when a small volume of blood is available has been developed. In this method two centrifugations of platelets layered on Ficoll-Uropoline cushion was applied. In the first step, platelet rich plasma layered over Ficoll-Uropoline solution was centrifuged (700 g, 30 min), bounded platelets recovered with the help of HEPES buffer, and centrifuged again (1000 g, 10 min) on Ficoll-Uropoline cushion. This rapid and simple procedure yielded platelets separated from von Willebrand factor, fibrinogen and albumin, as judged by bioassay and measurement of radiolabeled plasma proteins. The obtained preparation of platelets was stable and the platelet function preserved for 3 h as evidenced by platelet aggregation and fibrinogen binding analysis. The procedure is inexpensive and convenient for small volumes of blood.