Mazur E M, Hoffman R, Chasis J, Marchesi S, Bruno E
Blood. 1981 Feb;57(2):277-86.
The development of a satisfactory in vitro assay system for human megakaryocyte colony forming progenitor cells has been delayed by the lack of a suitable marker for cells of human megakaryocyte lineage. For this purpose we raised an antiserum directed against a purified human platelet glycoprotein preparation. In conjunction with indirect immunofluorescent staining of human bone marrow, this antiserum labeled only platelets, megakaryocytes, and an infrequent population of small mononuclear cells. These small mononuclear cells, not otherwise identifiable as members of the megakaryocyte series, constituted 22.9% of the total fluorescein positive nucleated bone marrow cells. This antiserum was also used to label colonies cultured from human peripheral blood mononuclear cells using a modified plasma clot technique. A mean of 123 fluorescein-labeled colonies were cloned per 10(6) mononuclear cells cultured. Granulocyte-macrophage and erythroid burst colonies did not label using this method. No augmentation of colony numbers was found with varying concentrations of erythropoietin, human embryonic kidney cell conditioned media (a source of thrombopoietin), or media conditioned by a human T lymphoblast cell line (a source of both colony stimulating and burst promoting activities). Immunofluorescent labeling for platelet glycoproteins is a convenient phenotypic marker for cells of human megakaryocyte lineage useful in the study of in vitro human megakaryocytopoiesis.
由于缺乏适用于人类巨核细胞系细胞的合适标志物，用于人类巨核细胞集落形成祖细胞的令人满意的体外检测系统的开发一直被推迟。为此，我们制备了一种针对纯化的人类血小板糖蛋白制剂的抗血清。结合人类骨髓的间接免疫荧光染色，这种抗血清仅标记血小板、巨核细胞以及少量的小单核细胞群体。这些小单核细胞无法通过其他方式被鉴定为巨核细胞系列的成员，占总荧光素阳性有核骨髓细胞的22.9%。这种抗血清还被用于标记使用改良血浆凝块技术从人类外周血单核细胞培养的集落。每培养10(6)个单核细胞平均可克隆出123个荧光素标记的集落。使用这种方法，粒细胞-巨噬细胞集落和红系爆式集落未被标记。在使用不同浓度的促红细胞生成素、人胚肾细胞条件培养基（一种血小板生成素来源）或人T淋巴母细胞系条件培养基（一种集落刺激和爆式促进活性来源）时，未发现集落数量增加。血小板糖蛋白的免疫荧光标记是人类巨核细胞系细胞的一种方便的表型标志物，可用于体外人类巨核细胞生成的研究。
