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人结肠癌细胞系Caco-2的上皮特性:电参数

Epithelial properties of human colonic carcinoma cell line Caco-2: electrical parameters.

作者信息

Grasset E, Pinto M, Dussaulx E, Zweibaum A, Desjeux J F

出版信息

Am J Physiol. 1984 Sep;247(3 Pt 1):C260-7. doi: 10.1152/ajpcell.1984.247.3.C260.

Abstract

Human colonic carcinoma Caco-2 cells grown in vitro undergo epithelial differentiation. Electrical measurements showed that they form resistant monolayers of polarized cells. On millipore filters, transepithelial electrical resistance (154 +/- 6.5 omega X cm2) was accompanied by a small potential difference (0.29 +/- 0.02 mV, serosal side positive) and by short-circuit current (1.9 +/- 0.14 microA X cm-2), both of which were ouabain sensitive. Micropuncture of domes formed on plastic supports under standard culture conditions revealed electrical polarity similar to that of filter-grown cells (0.8 +/- 0.2 mV, serosal side positive) combined with a highly negative cytoplasm (-57 +/- 1 mV) and very marked cell asymmetry (76% of total electrical cell resistance was located in the mucosal membrane). These parameters were not affected by the diuretic amiloride nor the hormone aldosterone, suggesting that sodium conductance is very limited in the mucosal membrane. Addition to the mucosal side of the ionophore nystatin or amphotericin B unmasked the possibility of high electrical transport activity. Electrical measurements made it possible to define the epithelial properties of Caco-2 cells, which may resemble those of colonic crypt or fetal cells. These measurements also confirmed that functional differentiation is homogeneous in Caco-2 cells. It is suggested that dome cell micropuncture may be useful in investigating the functional properties of other dome-forming cell lines.

摘要

体外培养的人结肠癌细胞Caco - 2会发生上皮分化。电生理测量表明,它们形成了极化细胞的抗性单层。在微孔滤膜上,跨上皮电阻(154±6.5Ω×cm²)伴随着较小的电位差(0.29±0.02 mV,浆膜侧为正)和短路电流(1.9±0.14μA×cm⁻²),这两者均对哇巴因敏感。在标准培养条件下,对塑料支架上形成的圆顶进行微穿刺,结果显示其电极性与滤膜培养的细胞相似(0.8±0.2 mV,浆膜侧为正),同时细胞质高度呈负(-57±1 mV),且细胞不对称性非常明显(总细胞电阻的76%位于黏膜膜)。这些参数不受利尿药氨氯吡咪或激素醛固酮的影响,这表明黏膜膜中的钠电导非常有限。在黏膜侧添加离子载体制霉菌素或两性霉素B揭示了高电转运活性的可能性。电生理测量使得定义Caco - 2细胞的上皮特性成为可能,这些特性可能类似于结肠隐窝或胎儿细胞的特性。这些测量还证实了Caco - 2细胞中的功能分化是均匀的。有人提出,圆顶细胞微穿刺可能有助于研究其他形成圆顶的细胞系的功能特性。

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