Chen Shuai, Einspanier Ralf, Schoen Jennifer
College of Life Science, Hebei University, Baoding, 071002, Hebei, People's Republic of China.
Institute of Reproductive Biology, Leibniz Institute for Farm Animal Biology, Wilhelm-Stahl-Allee 2, 18196, Dummerstorf, Germany.
Histochem Cell Biol. 2015 Nov;144(5):509-15. doi: 10.1007/s00418-015-1351-1. Epub 2015 Jul 31.
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*cm(2). 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.
在多孔滤器上培养输卵管上皮细胞可促进其呈现类似体内的形态和功能。然而,由于滤器材料的光学特性以及细胞的柱状形状,通过光学显微镜很难评估细胞质量。在本研究中,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置于四种不同类型的培养基中培养3周和6周以获得不同的培养质量,在处理样本进行组织学分析之前测量TEER。使用形态学标准(纤毛的存在、汇合度和细胞极性)对培养质量进行评分。我们还分析了在完全分化的常规培养物(生物学变异)以及因激素刺激导致细胞高度改变的培养物中,细胞高度(作为顶 - 基极化的指标)与TEER之间的相关性。完全分化的培养物的TEER在500至1100 Ω*cm²之间。在这个定义范围内呈现TEER值的培养物中,只有5%质量较差。亚分化的培养物显示TEER要么非常低要么过高。我们发现,在分化良好的培养物(常规培养组和激素刺激组)中,TEER与上皮高度之间存在高度显著(P < 0.0001)的负线性相关性。这可能表明紧密连接组装与上皮顶 - 基极化之间存在相互作用。总之,TEER是一个直接的质量指标,可常规用于监测体外输卵管上皮细胞的分化状态。