Dvorak A M, Massey W, Warner J, Kissell S, Kagey-Sobotka A, Lichtenstein L M
Department of Pathology, Beth Israel Hospital, Boston, MA 02215.
Blood. 1991 Feb 1;77(3):569-78.
Isolated human skin mast cells (HSMC) were prepared and cultured overnight before functional and electron microscopic studies. Mast cell suspensions were examined after stimulation with anti-IgE to produce anaphylactic degranulation or examined in buffer-incubated controls. Histamine release was measured in replicate samples. Control, isolated HSMC studied by electron microscopy were well preserved and fully granulated. Although all granule patterns reported for human mast cells were found, crystal granules were the most prevalent, as is true for HSMC in situ. Individual mast cells containing both crystal and scroll granules occurred. Lipid bodies were rare, as in HSMC in situ. Control, isolated mast cells did not express granule changes associated with either piecemeal degranulation or recovery during wound healing in situ; nor were morphologic changes of anaphylactic degranulation present. Spontaneous histamine release was 0% in control samples. Anaphylactic degranulation of isolated HSMC was accompanied by 24% maximum histamine release and characteristically showed extrusion of altered, membrane-free granules through multiple pores in the plasma membrane to the exterior of the cell. Other morphologic aspects of anaphylactic degranulation, as expressed in isolated human lung mast cells, were also present. These events included granule swelling, fusion, alteration of matrix contents, degranulation channel formation, pore formation, and shedding of granules, membranes, and surface processes. The ultrastructural morphology of isolated HSMC and their IgE-mediated degranulation shows some differences from similar studies of isolated human lung mast cells and of human lung and gut mast cells in biopsy samples. These differences include crystal granules as the predominant granule pattern, minor numbers of lipid bodies, and extrusion of granules during anaphylactic degranulation as characteristic for HSMC. By contrast, isolated human lung and gut mast cells have more scroll granules and particle granules, respectively, and more lipid bodies. In isolated human lung mast cells, anaphylactic degranulation is almost exclusively an intracellular fusion event characterized by the formation of complex degranulation channels within which altered granule matrix materials solubilize. In addition to morphologic differences between mast cells of skin, lung, or gut origin, functional differences have also been reported among mast cells of these organs. The ultrastructural morphology of isolated HSMC is identical to that of skin mast cells in biopsy samples, thereby validating the usefulness of this new source of HSMC for correlative functional and morphologic studies.
分离出的人皮肤肥大细胞（HSMC）在进行功能和电子显微镜研究前制备并培养过夜。用抗IgE刺激肥大细胞悬液以产生过敏反应性脱颗粒，或在缓冲液孵育的对照中进行检测。组胺释放量在重复样本中进行测定。对照样本中，通过电子显微镜研究的分离出的HSMC保存良好且颗粒饱满。虽然发现了所有报道的人肥大细胞的颗粒模式，但晶体颗粒最为普遍，原位的HSMC也是如此。同时存在含有晶体颗粒和卷轴颗粒的单个肥大细胞。脂体很少，原位的HSMC也是如此。对照样本中，分离出的肥大细胞未表现出与原位伤口愈合过程中的逐片脱颗粒或恢复相关的颗粒变化；也未出现过敏反应性脱颗粒的形态学变化。对照样本中组胺的自发释放率为0%。分离出的HSMC的过敏反应性脱颗粒伴随着最大24%的组胺释放，其特征是改变的、无膜颗粒通过质膜上的多个孔挤出到细胞外。在分离出的人肺肥大细胞中表现出的过敏反应性脱颗粒的其他形态学特征也存在。这些事件包括颗粒肿胀、融合、基质内容物改变、脱颗粒通道形成、孔形成以及颗粒、膜和表面突起的脱落。分离出的HSMC的超微结构形态及其IgE介导的脱颗粒与对分离出的人肺肥大细胞以及活检样本中的人肺和肠道肥大细胞的类似研究存在一些差异。这些差异包括晶体颗粒作为主要颗粒模式、少量脂体以及过敏反应性脱颗粒过程中颗粒的挤出是HSMC的特征。相比之下，分离出的人肺和肠道肥大细胞分别有更多的卷轴颗粒和颗粒状颗粒，以及更多的脂体。在分离出的人肺肥大细胞中，过敏反应性脱颗粒几乎完全是一种细胞内融合事件，其特征是形成复杂的脱颗粒通道，在通道内改变的颗粒基质物质溶解。除了皮肤、肺或肠道来源的肥大细胞之间的形态学差异外，这些器官的肥大细胞之间也报道了功能差异。分离出的HSMC的超微结构形态与活检样本中皮肤肥大细胞的形态相同，从而验证了这种新来源的HSMC用于相关功能和形态学研究的实用性。
