Clonal growth of human megakaryocyte progenitors in serum-free cultures: effect of recombinant human interleukin 3.
Clonal growth of human megakaryocyte progenitors in serum-free cultures: effect of recombinant human interleukin 3.
复制标题
人巨核细胞祖细胞在无血清培养物中的克隆生长：重组人白细胞介素 3 的作用。
作者：
M. Teramura;J. Katahira;S. Hoshino;T. Motoji;K. Oshimi;H. Mizoguchi
在含有去离子牛血清白蛋白、铁饱和转铁蛋白、2-巯基乙醇和重组人白细胞介素3（IL-3）的无血清琼脂培养基中观察到人巨核细胞集落形成。采用抗血小板糖蛋白IIb/IIIa单克隆抗体，通过碱性磷酸酶抗碱性磷酸酶技术原位鉴定巨核细胞集落。集落数量在第14天达到峰值，平均每2 × 10（5）个骨髓单核细胞有47个巨核细胞集落（范围23-104）。重组人IL-3刺激巨核祖细胞的生长。使用非贴壁、T细胞耗尽的单核细胞获得了类似的结果。这些发现表明，IL-3直接刺激巨核祖细胞的生长，而不激活辅助细胞，或不需要血清或血浆中存在的因子。该无血清培养体系可用于研究纯化的天然和重组生物调节剂对人巨核祖细胞的作用。
Human megakaryocyte colony formation was observed in serum-free agar cultures with a medium containing deionized bovine serum albumin, iron-saturated transferrin, 2-mercaptoethanol, and recombinant human interleukin 3 (IL-3). Megakaryocyte colonies were identified in situ by the alkaline phosphatase anti-alkaline phosphatase technique, using monoclonal antibody against platelet glycoprotein IIb/IIIa. Colony numbers reached a peak at day 14, with a mean of 47 megakaryocyte colonies (range 23-104) per 2 x 10(5) bone marrow mononuclear cells. Recombinant human IL-3 stimulated the growth of megakaryocyte progenitors. Similar results were obtained using nonadherent, T-cell-depleted mononuclear cells. These findings suggest that IL-3 stimulates the growth of megakaryocyte progenitors directly without activation of accessory cells, or the requirement of factors present in the serum or plasma. This serum-free culture system may be useful for studying the effects of purified natural and recombinant biological regulators on human megakaryocyte progenitors.