Nunthanasup N.Ketprasit N.Noulsri E.Palasuwan A.Combes V.Kulkeaw K.Palasuwan D.Mahidol University2024-02-082024-02-082023-12-01Scientific Reports Vol.13 No.1 (2023)https://repository.li.mahidol.ac.th/handle/123456789/95824The use of megakaryoblastic leukemia MEG-01 cells can help reveal the mechanisms of thrombopoiesis. However, conventional in vitro activation of platelet release from MEG-01 cells requires thrombopoietin, which is costly. Here, we aim to develop a more straightforward and affordable method. Synchronization of the MEG-01 cells was initially performed using serum-free culture, followed by spontaneous cell differentiation in the presence of serum. Different stages of megakaryoblast differentiation were classified based on cell morphology, DNA content, and cell cycle. The MEG-01 cells released platelet-like particles at a level comparable to that of the thrombopoietin-activated MEG-01 cells. The platelet-like particles were distinguishable from PLP-derived extracellular vesicles and could express P-selectin following ADP activation. Importantly, the platelet-like particles induced fibrin clotting in vitro using platelet-poor plasma. Therefore, this thrombopoietin-independent cell synchronization method is an effective and straightforward method for studying megakaryopoiesis and thrombopoiesis.MultidisciplinaryArticleSCOPUS10.1038/s41598-023-50111-62-s2.0-851802510752045232238110522