Measurements of transepithelial electrical resistance (TEER) are affected by junctional length in immature epithelial monolayers.
Measurements of transepithelial electrical resistance (TEER) are affected by junctional length in immature epithelial monolayers.
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DOI：
10.1007/s00418-021-02026-4
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发表时间：
2021-12
影响因子：
2.3
通讯作者：
Michael M
中科院分区：
文献类型：
--
作者：
Felix K;Tobias S;Jan H;Nicolas S;Michael M
跨上皮电阻（TEER）的测量是一种用于判定上皮细胞单层屏障完整性的常用技术。然而，值得注意的是，在类似条件下培养的相似细胞类型，其绝对TEER值存在极大的异质性。基于先前的观察，我们假设绝对TEER测量值的异质性不仅可以用连接蛋白的成熟来解释，还可以用单层内细胞连接绝对长度的动态变化来解释。因此，我们分析了Caco2细胞上皮细胞单层在分化过程中的TEER，特别关注连接复合体的变化以及单层内整体细胞形态的变化。我们发现，在上皮Caco2单层中，TEER在细胞达到汇合之前不断上升，随后下降一段时间，之后在连接分化过程中又进一步上升。相比之下，在不同时间点测量的大分子通透性，即4千道尔顿异硫氰酸荧光素（FITC） - 葡聚糖跨单层的通量，在此期间持续下降。详细分析表明，这一现象可以用分化过程中单层内细胞边界处连接长度的改变来解释。总之，这些观察结果证实，细胞数量的变化以及连接长度的随之增加对TEER值有至关重要的影响，尤其是在连接尚未成熟的早期汇合阶段。
The measurement of transepithelial electrical resistance (TEER) is a common technique to determine the barrier integrity of epithelial cell monolayers. However, it is remarkable that absolute TEER values of similar cell types cultured under comparable conditions show an immense heterogeneity. Based on previous observations, we hypothesized that the heterogeneity of absolute TEER measurements can not only be explained by maturation of junctional proteins but rather by dynamics in the absolute length of cell junctions within monolayers. Therefore, we analyzed TEER in epithelial cell monolayers of Caco2 cells during their differentiation, with special emphasis on both changes in the junctional complex and overall cell morphology within monolayers. We found that in epithelial Caco2 monolayers TEER increased until confluency, then decreased for some time, which was then followed by an additional increase during junctional differentiation. In contrast, permeability of macromolecules measured at different time points as 4 kDA fluorescein isothiocyanate (FITC)-dextran flux across monolayers steadily decreased during this time. Detailed analysis suggested that this observation could be explained by alterations of junctional length along the cell borders within monolayers during differentiation. In conclusion, these observations confirmed that changes in cell numbers and consecutive increase of junctional length have a critical impact on TEER values, especially at stages of early confluency when junctions are immature.
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