Ziomek C A, Schulman S, Edidin M
J Cell Biol. 1980 Sep;86(3):849-57. doi: 10.1083/jcb.86.3.849.
Single mouse intestinal epithelial cells (IEC) may be isolated by the use of a combination of methods used for the isolation of IEC from other species. Isolated cells remain viable for several hours. The membrane integral enzymes alkaline phosphatase and leucine aminopeptidase of isolated IEC are localized to the brush borders of IEC in tissue and in most newly isolated IEC. With time, both enzymes are found distributed over the entire cell surface. Redistribution appears to occur by diffusion in the plane of the membrane. It is slowed, but not blocked, if cells are maintained at 0 degrees C instead of at 37 degrees C, and it is not blocked by fixation in 0.5-3% paraformaldehyde. Drugs that alter cell membrane potential or that affect cell levels of ATP enhance the rate of redistribution of the enzymes.
通过使用从其他物种中分离肠上皮细胞(IEC)的多种方法的组合,可以分离出单个小鼠肠上皮细胞。分离出的细胞可存活数小时。分离出的IEC的膜整合酶碱性磷酸酶和亮氨酸氨肽酶在组织中和大多数新分离出的IEC中定位于IEC的刷状缘。随着时间的推移,两种酶都分布在整个细胞表面。重新分布似乎是通过在膜平面内的扩散发生的。如果将细胞维持在0℃而不是37℃,重新分布会减慢但不会被阻断,并且在0.5-3%多聚甲醛中固定也不会被阻断。改变细胞膜电位或影响细胞ATP水平的药物会提高酶的重新分布速率。