title: An efficient method for isolation of representative and contamination-free population of blood platelets for proteomic studies.
abstract: To date, there has been no ideal method for blood platelet isolation which allows one to obtain a preparation devoid of contaminations, reflecting the activation status and morphological features of circulating platelets. To address these requirements, we have developed a method which combines the continuous density gradient centrifugation with washing from PGI2-supplemented platelet-rich plasma (PRP). We have assessed the degree of erythrocyte and leukocyte contamination, recovery of platelets, morphological features, activation status, and reactivity of isolated platelets. Using our protocol, we were able to get a preparation free from contaminations, representing well the platelet population prior to the isolation in terms of size and activity. Besides this, we have obtained approximately 2 times more platelets from the same volume of blood compared to the most widely used method. From 10 ml of whole citrated blood we were able to get on average 2.7 mg of platelet-derived protein. The method of platelet isolation presented in this paper can be successfully applied to tests requiring very pure platelets, reflecting the circulating platelet state, from a small volume of blood.
authors: a Department of Medical Laboratory Diagnostics , Wroclaw Medical University , Wroclaw , Poland.
authors: Aneta
authors: A
authors: Wrzyszcz
authors: b Department of Laboratory Diagnostics , Lower Silesian Oncology Center , Wroclaw , Poland.
authors: Joanna
authors: J
authors: Urbaniak
authors: a Department of Medical Laboratory Diagnostics , Wroclaw Medical University , Wroclaw , Poland.
authors: Agnieszka
authors: A
authors: Sapa
authors: a Department of Medical Laboratory Diagnostics , Wroclaw Medical University , Wroclaw , Poland.
authors: Mieczysław
authors: M
authors: Woźniak
mesh_terms: D015415:Biomarkers; D001792:Blood Platelets / Q000378:metabolism*; D002469:Cell Separation / Q000379:methods* / Q000592:standards; D002499:Centrifugation, Density Gradient / Q000379:methods; D005434:Flow Cytometry; D006801:Humans; D063847:Mean Platelet Volume; D015539:Platelet Activation; D010974:Platelet Aggregation; D010976:Platelet Count; D010979:Platelet Function Tests; D020543:Proteome; D040901:Proteomics / Q000379:methods
chemical_list: D015415:Biomarkers; D020543:Proteome
keywords: Density gradient centrifugation; platelet isolation; platelet proteomics; platelets