Department of Oral Biology, School of Dental Medicine, University at Buffalo, The State University of New York, Buffalo, New York 14214-3092, USA.
Tissue Eng Part C Methods. 2010 Oct;16(5):1135-44. doi: 10.1089/ten.TEC.2009.0438.
The use of polarized salivary gland cell monolayers has contributed to our understanding of salivary gland physiology. However, these cell models are not representative of glandular epithelium in vivo, and, therefore, are not ideal for investigating salivary epithelial functions. The current study has developed a three-dimensional (3D) cell culture model for rat Par-C10 parotid gland cells that forms differentiated acinar-like spheres on Matrigel. These 3D Par-C10 acinar-like spheres display characteristics similar to differentiated acini in salivary glands, including cell polarization, tight junction (TJ) formation required to maintain transepithelial potential difference, basolateral expression of aquaporin-3 and Na+/K+/2Cl- cotransporter-1, and responsiveness to the muscarinic receptor agonist carbachol that is decreased by the anion channel blocker diphenylamine-2-carboxylic acid or chloride replacement with gluconate. Incubation of the spheres in the hypertonic medium increased the expression level of the water channel aquaporin-5. Further, the proinflammatory cytokines tumor necrosis factor-alpha and interferon-gamma induced alterations in TJ integrity in the acinar-like spheres without affecting individual cell viability, suggesting that cytokines may affect salivary gland function by altering TJ integrity. Thus, 3D Par-C10 acinar-like spheres represent a novel in vitro model to study physiological and pathophysiological functions of differentiated acini.
极化唾液腺细胞单层的使用有助于我们理解唾液腺生理学。然而,这些细胞模型不能代表体内的腺上皮,因此不适合研究唾液腺上皮功能。本研究开发了一种用于大鼠 Par-C10 腮腺细胞的三维(3D)细胞培养模型,该模型在 Matrigel 上形成分化的腺样球体。这些 3D Par-C10 腺样球体显示出与唾液腺中分化的腺泡相似的特征,包括细胞极化、维持跨上皮电位差所需的紧密连接(TJ)形成、水通道蛋白-3 和 Na+/K+/2Cl-共转运蛋白-1 的基底外侧表达,以及对毒蕈碱受体激动剂 carbachol 的反应性,该反应性被阴离子通道阻断剂二苯甲酰胺-2-羧酸或用葡萄糖酸盐替代氯离子所降低。将球体在高渗培养基中孵育会增加水通道 aquaporin-5 的表达水平。此外,促炎细胞因子肿瘤坏死因子-α和干扰素-γ诱导腺样球体中 TJ 完整性的改变,而不影响单个细胞活力,这表明细胞因子可能通过改变 TJ 完整性来影响唾液腺功能。因此,3D Par-C10 腺样球体代表了研究分化腺泡生理和病理生理功能的新型体外模型。