Zucker-Franklin D, Grusky G
J Cell Biol. 1974 Dec;63(3):855-63. doi: 10.1083/jcb.63.3.855.
The ultrastructure of granulocyte colonies derived from normal human peripheral blood leukocytes cultured in semisolid media has been studied by a new method developed for this purpose. Fixation, dehydration, and embedding of the whole content of the Petri dish resulted in a block of Epon containing colonies made up of cells with the spatial orientation of those observed in living cultures. This permitted serial sectioning through entire colonies. Cell maturation in vitro appeared to parallel that of normal marrow. However, even the most mature cells retained cytoplasmic characteristics of more immature cells. This was particularly true for eosinophils which only rarely possessed granules with electron-dense crystalline "cores," a feature typical for mature eosinophils. In addition to the normal-appearing hematopoietic cells found within colonies, very large round or spindle-shaped cells were present between colonies and firmly attached to the bottom of the culture dish. Although the histochemical and functional characterization of these cells awaits further study, it is suggested that they are related to histiocytes or macrophages. The technique described here should prove valuable in studies of the development, differentiation, and interaction of many types of cells.
利用为此目的开发的一种新方法，对在半固体培养基中培养的正常人外周血白细胞来源的粒细胞集落的超微结构进行了研究。对培养皿中的全部内容物进行固定、脱水和包埋，得到一块含有集落的环氧树脂包埋块，集落由细胞组成，这些细胞具有在活培养物中观察到的空间取向。这使得能够对整个集落进行连续切片。体外细胞成熟似乎与正常骨髓中的成熟情况相似。然而，即使是最成熟的细胞也保留了更多未成熟细胞的细胞质特征。嗜酸性粒细胞尤其如此，它们很少具有带有电子致密结晶“核心”的颗粒，而这是成熟嗜酸性粒细胞的典型特征。除了在集落中发现的外观正常的造血细胞外，在集落之间还存在非常大的圆形或梭形细胞，它们牢固地附着在培养皿底部。尽管这些细胞的组织化学和功能特性有待进一步研究，但有人认为它们与组织细胞或巨噬细胞有关。这里描述的技术在研究多种细胞的发育、分化和相互作用方面应该是有价值的。
