Department of Veterinary Medical Sciences, University of Bologna, Bologna, Italy.
Health Sciences and Technologies-Interdepartmental Center for Industrial Research (CIRI-SDV), Alma Mater Studiorum - University of Bologna, Bologna, Italy.
Methods Mol Biol. 2024;2749:151-164. doi: 10.1007/978-1-0716-3609-1_14.
A major feature of epithelial and endothelial cells is the creation of biological barriers able to protect the body against stressors that could compromise homeostasis. The ability to characterize biological barriers in vitro is an important study tool especially used for the intestinal barrier, the blood-brain barrier, and the lung barrier. The strength and integrity of biological barriers may be assessed by the measurement of the transepithelial/transendothelial electrical resistance (TEER) that reflects the ionic conductance of the paracellular pathway. The TEER measurement is a quantitative, non-invasive, highly useful, and representative method that must be strictly standardized. Here we describe a quantitative protocol to assess the mammary epithelial barrier integrity by combining the TEER measurement with a test for studying the passage of the sodium fluorescein, that is, a hydrophilic paracellular marker. Being the swine species an excellent translational model, primary cultures of mammary epithelial cells, isolated from hybrid pig tissue collected at slaughterhouse, are used.
上皮细胞和内皮细胞的一个主要特征是形成生物屏障,能够保护身体免受可能破坏体内平衡的应激源的影响。在体外表征生物屏障的能力是一种重要的研究工具,特别是用于肠道屏障、血脑屏障和肺屏障。生物屏障的强度和完整性可以通过测量跨上皮/跨内皮电阻 (TEER) 来评估,TEER 反映了细胞旁途径的离子电导率。TEER 测量是一种定量、非侵入性、非常有用且具有代表性的方法,必须严格标准化。在这里,我们描述了一种通过结合 TEER 测量和用于研究荧光素钠通透性的测试来评估乳腺上皮屏障完整性的定量方案,即亲水性细胞旁标记物。由于猪种是一种出色的转化模型,因此使用从屠宰场收集的杂种猪组织中分离的原代乳腺上皮细胞进行培养。