Grubb B R, Schiretz F R, Boucher R C
Cystic Fibrosis/Pulmonary Research and Treatment Center, University of North Carolina, Chapel Hill 27599-7248, USA.
Am J Physiol. 1997 Jul;273(1 Pt 1):C21-9. doi: 10.1152/ajpcell.1997.273.1.C21.
Airway epithelia are thought to play an important role in maintaining the depth (volume) and composition of airway surface liquid (ASL). However, due to the difficulty in measuring airway epithelial volume flow (Jv) and ASL composition, our knowledge of ASL homeostasis is limited. We have developed a permeable tubular culture system (biofiber) suitable for growing airway epithelia on the biofiber luminal surface, which allows measurements of bioelectric properties and Jv. Canine tracheal and bronchial epithelia readily attach, grow to confluence, and develop an electrical potential difference (-10 to -40 mV) across the biofiber. Using a six-hormone-supplemented medium, we detected a significant basal absorptive Jv across both the tracheal cells (0.65 +/- 0.08 microliter.cm-2.h-1) and bronchial cells (2.21 +/- 0.42 microliters.cm-2.h-1), which was significantly reduced by amiloride. Forskolin stimulated a net secretory Jv in tracheal biofibers (-0.56 +/- 0.19 microliter.h-1.cm-2) only. When the culture medium was supplemented with cholera toxin (CT), the basal absorptive Jv was significantly reduced in the bronchial biofibers and the tracheal biofibers exhibited net secretion. The forskolin-stimulated secretory Jv in the tracheal biofibers was significantly greater in the presence of CT than in its absence (-1.30 +/- 0.29 microliters.h-1.cm-2), whereas bronchial biofibers exhibited no significant Jv response to forskolin. We conclude that the Jv measured in tubular culture is highly dependent on the region from which the cells originated as well as the composition of the culture medium. Use of the biofiber culture system to study airway epithelia should give further insight into factors regulating Jv and composition of ASL.
气道上皮细胞被认为在维持气道表面液体(ASL)的深度(体积)和成分方面发挥着重要作用。然而,由于测量气道上皮细胞体积流量(Jv)和ASL成分存在困难,我们对ASL稳态的了解有限。我们开发了一种可渗透的管状培养系统(生物纤维),适合气道上皮细胞在生物纤维管腔表面生长,这使得能够测量生物电特性和Jv。犬气管和支气管上皮细胞很容易附着、生长至汇合,并在生物纤维上形成跨膜电位差(-10至-40 mV)。使用添加六种激素的培养基,我们检测到气管细胞(0.65±0.08微升·厘米-2·小时-1)和支气管细胞(2.21±0.42微升·厘米-2·小时-1)均存在显著的基础吸收性Jv,而氨氯吡咪可显著降低该Jv。福斯可林仅刺激气管生物纤维产生净分泌性Jv(-0.56±0.19微升·小时-1·厘米-2)。当培养基添加霍乱毒素(CT)时,支气管生物纤维的基础吸收性Jv显著降低,气管生物纤维则表现出净分泌。在存在CT的情况下,福斯可林刺激气管生物纤维产生的分泌性Jv显著大于不存在CT时(-1.30±0.29微升·小时-1·厘米-2),而支气管生物纤维对福斯可林未表现出显著的Jv反应。我们得出结论,在管状培养中测量的Jv高度依赖于细胞来源的区域以及培养基的成分。使用生物纤维培养系统研究气道上皮细胞应能进一步深入了解调节Jv和ASL成分的因素。