I use 9 ml of venous blood is drawn out in sterile containers, containing 1 ml of CPDA-1 (dissolve 2.99 citric acid anhydrous, 26.3 g sodium citrate dihydrate, 2.22 g sodium phosphate monobasic monohydrate, 31.9 g dextrose monohydrate, 0.275 g adenine and sterile dH2O sufficient to make 1000 ml mix and then filter and sterilize) that binds calcium and prevents blood clotting with no known interference to platelet function. The tube is gently sloped several times to mix the blood with the anticoagulant solution. A Double-Spin method (PRP-DS) at R.T. is use that provide after a first 150-200 g centrifugation (1500 rpm) for 5 minutes, the top layer of plasma is separated and centrifuged a second time (twenty minutes at 1000-2000 g [6300 rpm]). Finally, superficial plasma layer (platelet poor plasma - PPP) is fully removed and store at 4 °C. The platelet pellet is suspended in specific buffers or in PPP according to experiment to be performed. The whole PRP separation procedure is performed under aseptic conditions.
What is the intended use of the PRP? By a tobacco company, hmm...
I ask because of James Smith's query/comment. B-D states that their ACD (yellow top) tubes are for in vitro use.
Although sterile, I can as yet get no information from them regarding substances other than ACD, such as pyrogens or silicone, that could be harmful if the platelets are then administered to an animal or human.
University of Mumbai
Consorci MAR Parc de Salut de Barcelona
Sergio Lisboa Machado
Federal University of Rio de Janeiro
Instituto Politécnico de Lisboa
Tabaka Mission Hospital
Johns Hopkins University
Hannover Medical School
Loyola University Medical Center
Philip Morris International