Coleman D L, Tuet I K, Widdicombe J H
Am J Physiol. 1984 Mar;246(3 Pt 1):C355-9. doi: 10.1152/ajpcell.1984.246.3.C355.
Epithelial cells from dog trachea, when grown in tissue culture, formed confluent monolayers 5-6 days after plating. After 8-10 days, domes [mean diam 356 +/- (SE) 53 micron] appeared in monolayers grown in collagen-coated flasks. When grown on polycarbonate filters coated with collagen, a detectable resistance (greater than 5 omega X cm2) and transepithelial potential difference (PD) (greater than 0.1 mV) developed 6 days after plating and increased to approximately 15 omega X cm2 and 15 mV at 10 days. Serosal ouabain (10(-4) M) abolished PD and short-circuit current (Isc). Luminal ouabain had no effect. Luminal amiloride (10(-4) M) and serosal bumetanide (10(-4) M) each decreased PD and Isc. However, a combination of both of these drugs did not abolish Isc. Isoproterenol (10(-5) M), dibutyryl adenosine 3',5'-cyclic monophosphate (10(-3) M), vasoactive intestinal peptide (10(-7) M), prostaglandin (PG) E2 (10(-5) M), PGF2 alpha (10(-5) M), and bradykinin (10(-5) M) each increased PD and Isc. Thus these monolayer cultures maintain electrical properties resembling those of the original tissue. This preparation may prove useful for the study of water and ion transport by airway epithelia.
犬气管上皮细胞在组织培养中生长时,接种后5 - 6天形成汇合的单层细胞。8 - 10天后,在胶原包被的培养瓶中生长的单层细胞中出现穹顶(平均直径356±(标准误)53微米)。当在涂有胶原的聚碳酸酯滤膜上生长时,接种后6天出现可检测到的电阻(大于5Ω×cm²)和跨上皮电位差(PD)(大于0.1 mV),并在10天时增加到约15Ω×cm²和15 mV。浆膜侧哇巴因(10⁻⁴ M)消除了PD和短路电流(Isc)。管腔侧哇巴因无作用。管腔侧氨氯地平(10⁻⁴ M)和浆膜侧布美他尼(10⁻⁴ M)均降低了PD和Isc。然而,这两种药物联合使用并未消除Isc。异丙肾上腺素(10⁻⁵ M)、二丁酰腺苷3',5'-环磷酸(10⁻³ M)、血管活性肠肽(10⁻⁷ M)、前列腺素(PG)E2(10⁻⁵ M)、PGF2α(10⁻⁵ M)和缓激肽(10⁻⁵ M)均增加了PD和Isc。因此,这些单层培养物保持了类似于原始组织的电特性。这种制备方法可能对研究气道上皮细胞的水和离子转运有用。