Program in Clinical Hematology Sciences, Department of Clinical Microscopy, Faculty of Allied Health Sciences, Chulalongkorn University, Bangkok, 10330, Thailand.
Oxidation in Red Cell Disorders Research Unit, Department of Clinical Microscopy, Faculty of Allied Health Sciences, Chulalongkorn University, Bangkok, 10330, Thailand.
Sci Rep. 2023 Dec 18;13(1):22553. doi: 10.1038/s41598-023-50111-6.
The 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.
使用巨核母细胞白血病 MEG-01 细胞有助于揭示血小板生成的机制。然而，常规的体外激活 MEG-01 细胞释放血小板需要血小板生成素，这是昂贵的。在这里，我们旨在开发一种更简单和更经济的方法。首先使用无血清培养对 MEG-01 细胞进行同步化，然后在有血清的情况下自发地进行细胞分化。根据细胞形态、DNA 含量和细胞周期，将巨核母细胞的不同分化阶段进行分类。MEG-01 细胞释放出与血小板生成素激活的 MEG-01 细胞相当水平的血小板样颗粒。这些血小板样颗粒与 PLP 衍生的细胞外囊泡不同，并且可以在 ADP 激活后表达 P-选择素。重要的是，血小板样颗粒在体外使用血小板减少血浆诱导纤维蛋白凝块。因此，这种不依赖血小板生成素的细胞同步化方法是研究巨核细胞生成和血小板生成的有效且简单的方法。
