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[离体肠上皮的网格电子显微镜研究]

[Grid electron microscopy studies of isolated intestinal epithelium].

作者信息

Böck P, Tillmann B, Osterkamp U

出版信息

Z Mikrosk Anat Forsch. 1980;94(6):1077-89.

PMID:6792808
Abstract

Four media are used to isolate enterocytes from the jejunum of the mouse (basic medium with either hyaluronidase, dispase, pronase, or EDTA). During incubation the intestinal wall is gently agitated. Layers of epithelial cells rather than single cells are removed from the mucosal surface by these procedures. Small groups of enterocytes and single cells can be obtained by further mechanical agitation, e.g. by repeatedly washing the isolated epithelia. Similar morphological findings (transmission and scanning electron microscopy) are obtained after using the various isolation media. Only with EDTA a dilatation of the endoplasmic reticulum is regularly seen. Isolation of the enterocytes starts from the tip of the intestinal villus. The straight apical surface becomes convex and the most lateral microvilli of the brush border desintegrate. The opening of the terminal bars can be observed. Surface differentiations on the lateral cell surfaces of isolated enterocytes become less obvious and disappear. This is discussed with respect to the so-called "lateral vacuoles" which appear in isolated enterocytes. Large spherical protrusions are developed at the basal surface of enterocytes. The cell membrane covering these protrusions is frequently found to be discontinuous. No other cell organelles than ribosomes are to be seen inside of these protrusions. Membrane-bound vesicles are also frequently seen. It is suggested that the basal spherical protrusions are casted off.

摘要

使用四种培养基从小鼠空肠中分离肠上皮细胞(含透明质酸酶、分散酶、链霉蛋白酶或乙二胺四乙酸的基础培养基)。孵育过程中轻轻搅动肠壁。通过这些操作从黏膜表面去除的是上皮细胞层而非单个细胞。通过进一步的机械搅动,例如反复冲洗分离出的上皮组织,可以获得小群的肠上皮细胞和单个细胞。使用各种分离培养基后可得到类似的形态学结果(透射电子显微镜和扫描电子显微镜观察)。仅使用乙二胺四乙酸时,内质网会经常出现扩张。肠上皮细胞的分离从肠绒毛尖端开始。笔直的顶端表面变得凸起,刷状缘最外侧的微绒毛解体。可以观察到闭锁小带的开口。分离出的肠上皮细胞侧面的表面分化变得不那么明显并消失。这与分离出的肠上皮细胞中出现的所谓“侧泡”有关。肠上皮细胞的基底表面会形成大的球形突起。覆盖这些突起的细胞膜经常被发现是不连续的。在这些突起内部除了核糖体看不到其他细胞器。膜结合囊泡也经常可见。有人认为基底球形突起会脱落。

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