Nagata Y, Nagahisa H, Aida Y, Okutomi K, Nagasawa T, Todokoro K
Tsukuba Life Science Center, Institute of Physical and Chemical Research (RIKEN), Ibaraki, Japan.
J Biol Chem. 1995 Aug 25;270(34):19673-5. doi: 10.1074/jbc.270.34.19673.
Thrombopoietin (Tpo) is a cytokine that specifically regulates megakaryocyte maturation and platelet production. Little is known about the molecular and cellular mechanism of the Tpo-induced megakaryocyte maturation process including polyploidization and platelet release. To study Tpo-induced megakaryocyte differentiation, a mouse cell line FD-TPO, which responds and grows with Tpo, was established from a interleukin-3-dependent hematopoietic progenitor cell line FDC-P2. The FD-TPO cells, expressing endogenous Tpo receptor, grew with Tpo in a dose-dependent manner. Further, Tpo stimulation dramatically induced expression of megakaryocyte/erythroid-specific transcription factors GATA-1 and NF-E2 in FD-TPO cells. Flow cytometry analysis demonstrated that expression of platelet-specific cell surface antigens including CD61 (GPIIIa) dramatically increased in Tpo-stimulated FD-TPO cells and that expression of myeloid-specific antigens, Gr-1 and Mac-1, decreased. Therefore, we concluded that the binding of Tpo to FD-TPO cells induces not only cell growth but also differentiation into mature megakaryocyte-like cells, and thus this cell line was found to be useful for the study of Tpo receptor-mediated growth and differentiation signals.
血小板生成素（Tpo）是一种特异性调节巨核细胞成熟和血小板生成的细胞因子。关于Tpo诱导的巨核细胞成熟过程（包括多倍体化和血小板释放）的分子和细胞机制，我们了解甚少。为了研究Tpo诱导的巨核细胞分化，我们从依赖白细胞介素-3的造血祖细胞系FDC-P2中建立了一种小鼠细胞系FD-TPO，该细胞系对Tpo有反应并能在Tpo存在的情况下生长。表达内源性Tpo受体的FD-TPO细胞在Tpo存在的情况下呈剂量依赖性生长。此外，Tpo刺激显著诱导了FD-TPO细胞中巨核细胞/红细胞特异性转录因子GATA-1和NF-E2的表达。流式细胞术分析表明，在Tpo刺激的FD-TPO细胞中，包括CD61（GPIIIa）在内的血小板特异性细胞表面抗原的表达显著增加，而髓系特异性抗原Gr-1和Mac-1的表达则下降。因此，我们得出结论，Tpo与FD-TPO细胞的结合不仅诱导细胞生长，还诱导其分化为成熟的类巨核细胞，因此发现该细胞系可用于研究Tpo受体介导的生长和分化信号。
