An efficient method for isolation of representative and contamination-free population of blood platelets for proteomic studies

• Published in: Platelets (Edinburgh) Vol. 28; no. 1; pp. 43 - 53
• Main Authors: Wrzyszcz, Aneta, Urbaniak, Joanna, Sapa, Agnieszka, Woźniak, Mieczysław
• Format: Journal Article
• Language: English
• Published: England Taylor & Francis 01.01.2017; Taylor & Francis Group
• Subjects: Biomarkers; Blood Platelets - metabolism; Cell Separation - methods; Cell Separation - standards; Centrifugation, Density Gradient - methods; Density gradient centrifugation; Flow Cytometry; Humans; Mean Platelet Volume; Platelet Activation; Platelet Aggregation; Platelet Count; Platelet Function Tests; platelet isolation; platelet proteomics; platelets; Proteome; Proteomics - methods; Density gradient centrifugation; platelet isolation; platelet proteomics; platelets
• ISSN: 0953-7104; 1369-1635
• DOI: 10.1080/09537104.2016.1209478

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 PGI 2 -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.