Choi E S, Nichol J L, Hokom M M, Hornkohl A C, Hunt P
AMGEN, Inc., Thousand Oaks, CA 91320.
Blood. 1995 Jan 15;85(2):402-13.
An in vitro culture system demonstrating the transitions from megakaryocyte progenitors to functional platelets is described. CD34-selected cells from normal human peripheral blood are cultured under conditions that promote megakaryocyte formation. After 8 to 11 days, enriched populations of mature megakaryocytes are replated under conditions that favor the development of proplatelets. Proplatelets express the platelet-specific proteins, glycoproteins Ib and IIb (GPIb and GPIIb), and fibrinogen and also contain microtubule coils equal in size to those found in plasma-derived platelets. In addition, proplatelets have ultrastructural features in common with plasma-derived platelets. Platelet-sized particles from the proplatelet culture supernatants are examined. Ultrastructurally, these particles are identical to plasma-derived platelets. Functionally, these culture-derived platelets aggregate in response to both thrombin and adenosine diphosphate (ADP) plus fibrinogen. This aggregation is specifically inhibited by the addition of a function-blocking anti-GPIIbIIIa antibody. Culture-derived platelets stimulated with agonists also express the activation-dependent antigens P-selectin and functional fibrinogen receptor. This is the first description of an in vitro culture system that sequentially demonstrates megakaryocyte growth, development, and platelet production.
本文描述了一种体外培养系统，该系统可展示从巨核细胞祖细胞到功能性血小板的转变过程。从正常人外周血中选取经CD34筛选的细胞，在促进巨核细胞形成的条件下进行培养。8至11天后，将富集的成熟巨核细胞群体重新接种到有利于前血小板发育的条件下。前血小板表达血小板特异性蛋白、糖蛋白Ib和IIb（GPIb和GPIIb）以及纤维蛋白原，并且还含有与血浆来源血小板中大小相同的微管盘绕结构。此外，前血小板具有与血浆来源血小板相同的超微结构特征。对来自前血小板培养上清液的血小板大小颗粒进行检查。在超微结构上，这些颗粒与血浆来源血小板相同。在功能上，这些培养来源的血小板对凝血酶和二磷酸腺苷（ADP）加纤维蛋白原均有聚集反应。加入功能阻断性抗GPIIbIIIa抗体可特异性抑制这种聚集。用激动剂刺激培养来源的血小板也会表达活化依赖性抗原P-选择素和功能性纤维蛋白原受体。这是首次对一种体外培养系统的描述，该系统可依次展示巨核细胞的生长、发育和血小板生成过程。
