Scanning electron microscopy of kidney cells in culture: surface features of polarized epithelia.
Scanning electron microscopy of kidney cells in culture: surface features of polarized epithelia.
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发表时间：
1986
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通讯作者：
J. Mcateer;G. S. Dougherty;K. Gardner;A. Evan
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作者：
J. Mcateer;G. S. Dougherty;K. Gardner;A. Evan
我们已经使用扫描电子显微镜（SEM）来检查肾上皮细胞系MDCK和LLC-PK 1的表面形态，以确定替代培养基质条件对细胞极性的影响。我们观察到，无论物理培养条件，细胞建立和保持极性，表示的顶端和基底表面的特征。然而，培养条件确实影响体外细胞极性的方向。MDCK细胞在胶原凝胶内生长，其中单个细胞表现出克隆生长以形成充满液体的上皮囊肿。MDCK-囊肿的细胞被极化，顶面朝向管腔，底面朝向周围的胶原凝胶。这种配置使得通过去除支持性胶原网格，通过SEM可以直接观察基底表面。MDCK囊肿的顶端表面排列着短的微绒毛。每个细胞都有一个单独的纤毛。相比之下，基表面有很少的附属物，虽然细胞边界是由交错的短突起。单层培养的LLC-PK 1细胞在其顶端表面具有孤纤毛和长微绒毛。观察到参与圆顶形成的细胞的基底表面仅具有稀疏的短而钝的突起。当LLC-PK 1细胞在静止悬浮培养物中或在非培养级塑料上的单层中培养时，它们形成细胞顶端面向周围培养基的包囊。这些细胞表现出可变的顶端形态。大的，高度膨胀的囊肿的细胞往往是衰减的，并有一个相对光滑的顶面。破裂的LLC-PK 1囊肿的细胞基底表面通常也是光滑的，没有突出的附属物。
We have used scanning electron microscopy (SEM) to examine the surface morphology of the renal epithelial cell lines MDCK and LLC-PKl to determine the influence of alternative culture substrate conditions on cell polarity. We observed that regardless of physical culture conditions, cells established and maintained polarity, expressed by the characteristics of apical and basal surfaces. Culture conditions did, however, influence the orientation of cell polarity in vitro. MDCK cells were grown within collagen gel, in which individual cells exhibited clonal growth to form fluid-filled epithelial cysts. The cells of MDCK-cysts were polarized with apical surface facing the lumen and basal surface against the surrounding collagen gel. This configuration made it possible to gain direct visual access, by SEM, to the basal surface by removing the supportive collagen lattice. The apical surface of MDCK-cysts was lined by short microvilli. Each cell possessed a solitary cilium. In comparison, the basal surface had few appendages, although cell boundaries were marked by interdigitating short processes. LLC-PKl cells in monolayer culture bore solitary cilia and long microvilli at their apical surface. The basal surface of cells involved in dome formation was observed to possess only a sparse population of short, blunt processes. When LLC-PKl cells were raised in stationary suspension culture or in monolayer atop non-culture grade plastic, they formed cysts with the cell apex facing the surrounding medium. These cells showed variable apical morphology. The cells of large, highly expanded cysts were often attenuated and had a relatively smooth apical surface. The basal surface of cells of fractured LLC-PKl cysts commonly was also smooth, without prominent appendages.