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Transepithelial electrical resistance (TEER): a functional parameter to monitor the quality of oviduct epithelial cells cultured on filter supports

机译:跨上皮电阻(TEER):一个功能参数用于监测培养在滤膜支架上的输卵管上皮细胞的质量

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摘要

Cultivation of oviduct epithelial cells on porous filters fosters in vivo-like morphology and functionality. However, due to the optical properties of the filter materials and the cells’ columnar shape, cell quality is hard to assess via light microscopy. In this study, we aim to evaluate transepithelial electrical resistance (TEER) measurement as a prognostic quality indicator for the cultivation of porcine oviduct epithelial cells (POEC). POEC were maintained in four different types of media for 3 and 6 w to achieve diverse culture qualities, and TEER was measured before processing samples for histology. Culture quality was scored using morphological criteria (presence of cilia, confluence and cell polarity). We furthermore analyzed the correlation between cellular height (as a measure of apical–basal polarization) and TEER in fully differentiated routine cultures (biological variation) and in cultures with altered cellular height due to hormonal stimulation. Fully differentiated cultures possessed a moderate TEER between 500 and 1100 Ω*cm2. Only 5 % of cultures which exhibited TEER values in this defined range had poor quality. Sub-differentiated cultures showed either very low or excessively high TEER. We unveiled a highly significant (P < 0.0001) negative linear correlation between TEER and epithelial height in well-differentiated cultures (both routine and hormone stimulated group). This may point toward the interaction between tight junction assembly and epithelial apical–basal polarization. In conclusion, TEER is a straightforward quality indicator which could be routinely used to monitor the differentiation status of oviduct epithelial cells in vitro.Electronic supplementary materialThe online version of this article (doi:10.1007/s00418-015-1351-1) contains supplementary material, which is available to authorized users.
机译:在多孔滤膜上培养输卵管上皮细胞可促进体内类似的形态和功能。但是,由于滤材的光学特性和细胞的柱状形状,很难通过光学显微镜评估细胞质量。在这项研究中,我们旨在评估跨上皮电阻(TEER)测量作为猪输卵管上皮细胞(POEC)培养的预后质量指标。将POEC分别在4种不同类型的培养基中分别维持3和6 w的质量以达到不同的培养质量,并在处理组织学样品之前测量TEER。使用形态学标准(纤毛的存在,融合和细胞极性)对培养质量进行评分。我们还分析了完全分化的常规培养物(生物学变异)和由于激素刺激而引起细胞高度改变的培养物中细胞高度(作为根尖-基础极化的量度)与TEER之间的相关性。完全分化的培养物的TEER在500至1100Ω* cm 2 之间。在此限定范围内显示TEER值的培养物中只有5%的质量较差。亚分​​化培养物显示TEER极低或过高。我们揭示了在高度分化的文化(常规和激素刺激组)中TEER与上皮高度之间的高度显着(P <0.0001)负线性相关。这可能指向紧密连接组件和上皮根尖-基底极化之间的相互作用。总之,TEER是一种直接的质量指标,可常规用于体外监测输卵管上皮细胞的分化状态。电子补充材料本文的在线版本(doi:10.1007 / s00418-015-1351-1)包含补充材料,可供授权用户使用。

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