MORPHOLOGY OF FIBROBLASTS IN COLLAGEN GELS - A STUDY USING 400 KEV ELECTRON-MICROSCOPY AND COMPUTER-GRAPHICS
MORPHOLOGY OF FIBROBLASTS IN COLLAGEN GELS - A STUDY USING 400 KEV ELECTRON-MICROSCOPY AND COMPUTER-GRAPHICS
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DOI：
10.1002/cm.970140308
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发表时间：
1989-01-01
影响因子：
--
通讯作者：
PEACHEY, LD
中科院分区：
文献类型：
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作者：
HEATH, JP;PEACHEY, LD
我们使用了400千伏的中压电子显微镜(IVEM)来观察在胶原凝胶中培养的成纤维细胞的厚片。在这些3D胶原晶格中，向外延伸并与胶原基质接触的狭长伪足延伸显示出一种复杂的地形，这是细胞在平面底物上移动时所没有的。因此，包围整个细胞过程的1至2微米厚的部分比薄的部分更能反映细胞和胶原之间相互作用的整体形态。用抗体和蛋白A-胶体金偶联物标记了一些细胞，以帮助在显微立体图中辨别细胞突起的形态。这些金颗粒为计算机辅助从倾斜序列IVEM图像重建膜形态提供了接近清晰的3D参考点。我们的结果证实，通过胶原蛋白晶格移动的细胞缺乏在玻璃上移动的细胞的良好铺展形态。它们通常相当细长，有几个长的分枝的前伪足。我们发现细胞体和主要的伪足突起是圆柱形的，正如人们对3D环境中细胞的预期，但在前进的伪足的前沿有一些小的扁平延伸，类似于玻璃上的细胞。这种相似性表明，片状基质是成纤维细胞在所有类型的底物上运动时使用的一种基本类型的突起结构。扁平的片状脂膜可能是细胞在胚胎形态发生过程中感知纤维状细胞外基质的部分机制。
We have used 400 kilovolt intermediate voltage electron microscopy (IVEM) to examine thick sections of fibroblasts cultured in collagen gels. In these 3D collagen lattices, the long, narrow pseudopodial extensions that extend out and make contact with the collagen matrix exhibit a complex topography not seen in the processes put out by cells moving on planar substrate. For this reason, sections 1 to 2 .mu.m thick that enclose a whole cell process are more informative of the overall morphology of the interaction between cells and the collagen than are thin sections. To aid the discrimination of topography of cell processes in stereo views of micrographs, some cells were labeled with antibodies and protein A-colloidal gold conjugates. The gold particles provided near clear 3D reference points for computer-aided reconstruction of membrane topography from tilt series of IVEM images. Our results confirm that cells that move through collagen lattices lack the well-spread morphology of their counterparts moving on glass. They are generally rather spindly with several long branching anterior pseudopodia. We found that the cell bodies and major pseudopodial processes were cylindrical, as one might expect of cells in a 3D environment, but at the leading edge of advancing pseudopodia there are small flat extensions similar to those seen in cells on glass. This similarity suggests that the lamellipodium is a basic type of protrusive structure used by fibroblasts during locomotion on all types of substratum. The flattened shape of lamellipodia may be part of the mechanism by which cells sense the oritentation of fibrillar extracellular matrices during embryonic morphogenesis.