Question 1: I would like to isolate platelets without activating them beforehand and also at the same time make sure that I can analyse them in non-aggregated forms (i.e. Individual platelets). Will that be possible, and if so are there particular protocols/methods I can refer to?
Question 2: What about achieving the above platelets isolation from blood samples of patients with disease associated with platelets activation and aggregation (e.g. heart disease patients)? Is there a way to ensure that the resultant isolate contains platelets that are not aggregated or adhered to each other or other particulates?
Thank you for taking time to read my question and for providing answers/advice!
For both questions, the answer is the same. Yes you can. The important thing is to anti-coagulate the blood.
We usually collect whole blood into acid- citrate-dextrose (ACD, 3.2%) sterile tubes. Depending on your read-out you can use this blood for flow cytometry, alternatively you can subsequently isolate the platelets.
Briefly, the whole blood needs to be first centrifuged at 150xg for 20min at room temperature (RT). The platelet-rich plasma (PRP) then needs to be removed and PGE1 was added to the PRP to prevent exogenous platelet activation. The PRP is then centrifuged at 400xg for 20min at RT. The platelet pellet was re-suspended in warmed (37oC) PIPES saline glucose (PSG).
More details can be found here:
PMID: 32573711
PMID: 31366617
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