Identification and characterization of the monoblast in mononuclear phagocyte colonies grown in vitro.
Identification and characterization of the monoblast in mononuclear phagocyte colonies grown in vitro.
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DOI：
10.1084/jem.142.5.1180
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发表时间：
1975-11-01
期刊：
影响因子：
--
通讯作者：
van Furth R
中科院分区：
文献类型：
--
作者：
Goud TJ;Schotte C;van Furth R
描述了一种在玻璃表面培养单核巨噬细胞集落的液体培养技术。这项有用而可靠的技术使研究未成熟的单核吞噬细胞成为可能。在单核吞噬细胞集落中，细胞在单层中彼此分开生长。在这些集落中可识别三种类型的细胞，即未分裂的巨噬细胞、增殖的原单核细胞和单核细胞。巨噬细胞和原单核细胞表现出其他方法所表现出的典型特征，而单核细胞只能用目前的液体培养方法来完全表征。这种增殖细胞([~3H]胸腺嘧啶核苷标记指数为92-96%)几乎呈圆形(直径10×10微米)，只有一小圈强嗜碱性的细胞质，几乎没有颗粒，细胞表面有一定程度的褶皱。以α-萘丁酸酯为底物的单核细胞对酯酶呈阳性反应(91%)，对过氧化物酶呈阳性反应(在过氧化物酶阳性菌落中占78%)，对溶菌酶呈阳性反应(43%)。该单核细胞能够吞噬葡聚糖硫酸盐(15-20%)和吞噬调理细菌(20%-30%)、乳胶颗粒(47%)和包被免疫球蛋白的红细胞(96%)。细胞表面存在免疫球蛋白G受体(94%)和补体受体(16%)。在这些方面，单核细胞具有单核吞噬细胞的典型特征，但其特性表明它是一种比原单核细胞更不成熟的细胞类型。根据这些标准和不同细胞类型在孵育和单个单核巨噬细胞集落发育过程中出现的顺序，单核细胞先于原单核细胞出现在集落中，从而得出单核细胞是原单核细胞的前体的结论。在这些培养物中也形成粒细胞集落，由成髓细胞、(亲)粒细胞、STAB和中性粒细胞组成。除了典型的紧密结构外，粒细胞本身在形态、细胞化学特征(α-萘丁酸酯酶阴性，N-乙酰-DL-丙氨基-1-萘酯阳性96%)、功能特征(胞饮指数13-21%；吞噬指数；调理细菌15-36%，乳胶颗粒10%，包被免疫球蛋白的红细胞0%)，以及极少量的IgG受体和缺乏补体受体等方面与单核吞噬细胞有很大的不同。根据这些标准，这些粒细胞很容易与单核巨噬细胞集落的未成熟细胞区分开来。本研究证实了单核巨噬细胞是一个独立的细胞系的结论，与粒细胞系完全不同，因为即使是迄今发现的最不成熟的细胞--单核细胞和成髓细胞--也有非常不同的特征。
A liquid culture technique for growing mononuclear phagocyte colonies on a glass surface is described. This useful and reliable technique made it possible to study immature mononuclear phagocytes. In the mononuclear phagocyte colonies the cells grow separate from each other in a single layer. Three types of cells are recognized in these colonies, namely nondividing macrophages, and proliferating promonocytes and monoblasts. The macrophage and the promonocyte exhibit the typical characteristics previously demonstrated by the other methods, whereas the monoblast could only be fully characterized by the present liquid culture method. This proliferating cell (labeling index with [3H]thymidine, 92-96%) is almost round (diameters, 10 X 10 mum), has only a small rim of strongly basophilic cytoplasm, almost devoid of granules, and shows a certain degree of ruffling of the cell surface. The monoblast is positive for esterase with alpha-naphthyl butyrate as substrate (91%), for peroxidase (78% in the peroxidase-positive colonies), and lysozyme (43%). The monoblast is able to pinocytize dextran sulphate (15-20%) and to phagocytize opsonized bacteria (20- 30%), latex particles (47%), and IgG-coated red cells (96%). IgG receptors (94%) and complement receptors (16%) are present at the cell surface. In these respects the monoblast has the typical characteristics of the mononuclear phagocytes, but its properties show it to be a more immature cell type than the promonocyte. On the basis of these criteria and the sequence of appearance of the different cell types during incubation and during the development of the individual mononuclear phagocyte colony, monoblasts being present before promonocytes appear in the colony, it is concluded that the monoblast is the precursor of the promonocyte. In these cultures granulocyte colonies are also formed, consisting of myeloblasts, (pro)myelocytes, stabs, and polymorphonuclear neutrophils. Besides the typically tight structure of this kind of colony, the granulocytic cells themselves are quite distinct from the mononuclear phagocytes by their morphology, cytochemical characteristics (e.g. all negative for esterase with alpha- naphthyl butyrate, but 96% positive with N-acetyl DL-alanyl 1- naphthylester), functional characteristics (pinocytic index 13-21%; phagocytic index; for opsonized bacteria 15-36%, for latex particles 10%, and for IgG-coated red cells 0%), and their very small number of IgG receptors and lack of complement receptors. On the basis of these criteria, these granulocytic cells are easily distinguished from the immature cells of the mononuclear phagocyte colonies. The present study confirms the conclusion that the mononuclear phagocytes are a separate cell line, quite distinct from the granulocytic series, since even the most immature cells so far identified--the monoblast and the myeloblast- -have quite different characteristics.