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

机译：一种有效的方法，用于分离出代表性和无污染的血小板血小板群进行蛋白质组学研究

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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.
机译：迄今为止，没有理想的血小板隔离方法，其允许人获得缺乏污染的制剂，反映循环血小板的激活状态和形态学特征。为了解决这些要求，我们开发了一种将连续密度梯度离心与从PGI2补充的富含血小板的血浆（PRP）的洗涤相结合的方法。我们评估了红细胞和白细胞污染的程度，血小板恢复，形态特征，激活状态和分离血小板的反应性。使用我们的协议，我们能够获得免于污染的准备，在尺寸和活动方面均衡血小板群体。除此之外，与最广泛使用的方法相比，我们从相同体积的血液中获得了大约2倍的血小板。从10毫升整个柑橘血液中，我们能够平均为2.7mg血小板衍生蛋白。本文呈现的血小板分离方法可以成功地应用于需要非常纯血小板的试验，从少量血液中反映循环血小板状态。

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