Yankaskas J R, Cotton C U, Knowles M R, Gatzy J T, Boucher R C
Am Rev Respir Dis. 1985 Dec;132(6):1281-7. doi: 10.1164/arrd.1985.132.6.1281.
The feasibility of culturing human nasal epithelial cells was tested by plating on permeable collagen matrices primary isolates of epithelial cells from normal (n = 21) and atopic (n = 5) subjects and from subjects with cystic fibrosis (CF) (n = 13). The cells were cultured in serum-free, hormone-supplemented medium for 5 days followed by maintenance in conditioned medium with 1% serum. A transepithelial electric potential difference (PD) exceeding 1.0 mV developed in most preparations (71%). The PD reached a peak by Day 10 and declined slowly thereafter. The maximal PD of CF epithelia (-32.4 +/- 5.8 mV, n = 9) on collagen matrices exceeded that of normal (-11.3 +/- 1.8 mV, n = 15) and atopic (-13.5 +/- 2.5, n = 5) epithelia. The maximal PD for both CF and normal cultures exceeded that of freshly excised preparations. This difference reflected higher transepithelial resistance of cultures as compared with intact preparations. The efficacy of amiloride (10(-4) M) to reduce the PD was greater in cultures of CF epithelia (-85 +/- 4.6%, n = 7) than it was in cultures of normal (-45.5 +/- 5.0%, n = 11) and atopic (-50.5 +/- 2.9%, n = 5) epithelia, whereas the hyperpolarization induced by replacement of Cl- in the apical bath was smaller for CF cultures (5.9 +/- 1.5 mV) than it was for normal (9.3 +/- 0.9 mV) and atopic (8.8 +/- 0.5 mV) cultures. We conclude that nasal epithelial cells will form high resistance barriers when cultured on collagen matrices.(ABSTRACT TRUNCATED AT 250 WORDS)
通过将来自正常受试者（n = 21）、特应性受试者（n = 5）和囊性纤维化（CF）受试者（n = 13）的上皮细胞原代分离物接种于可渗透胶原基质上，测试了培养人鼻上皮细胞的可行性。细胞在无血清、添加激素的培养基中培养5天，随后在含1%血清的条件培养基中维持培养。大多数制剂（71%）中出现了超过1.0 mV的跨上皮电位差（PD）。PD在第10天达到峰值，此后缓慢下降。胶原基质上CF上皮细胞的最大PD（-32.4±5.8 mV，n = 9）超过正常上皮细胞（-11.3±1.8 mV，n = 15）和特应性上皮细胞（-13.5±2.5，n = 5）。CF和正常培养物的最大PD均超过新鲜切除制剂。这种差异反映出与完整制剂相比，培养物的跨上皮电阻更高。氨氯地平（10⁻⁴ M）降低CF上皮细胞培养物中PD的效果（-85±4.6%，n = 7）大于正常上皮细胞培养物（-45.5±5.0%，n = 11）和特应性上皮细胞培养物（-50.5±2.9%，n = 5），而顶端浴中Cl⁻替代诱导的CF培养物超极化（5.9±1.5 mV）小于正常培养物（9.3±0.9 mV）和特应性培养物（8.8±0.5 mV）。我们得出结论，鼻上皮细胞在胶原基质上培养时会形成高电阻屏障。（摘要截短于250字）
