Culture conditions and treatments affect Caco-2 characteristics and particle uptake
Culture conditions and treatments affect Caco-2 characteristics and particle uptake
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DOI：
10.1016/j.ijpharm.2009.11.027
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发表时间：
2010-03-15
影响因子：
5.8
通讯作者：
Carr, K. E.
中科院分区：
文献类型：
--
作者：
Moyes, S. M.;Morris, J. F.;Carr, K. E.
小肠微粒通过细胞旁途径的摄取与口服药物递送和环境污染有关。体外研究使用乳胶微粒穿过汇合的 Caco-2 细胞上皮。本文探讨了培养条件对跨上皮抵抗（TER）的影响；共聚焦显微镜的细胞尺寸；和上皮下方的颗粒数量。研究的变量包括初始 TER 水平；多次 TER 测量；媒介的参与；细胞来源；并用乙醇或一定温度进行预处理。暴露于2μm乳胶颗粒5-120分钟后收集数据：假手术组暴露于预处理但不暴露于颗粒。结果强调了非常精确控制实验环境的重要性：确认仅暴露于颗粒的组中 TER 连续增加/减少的模式，并显示细胞尺寸随之增加。更大的颗粒摄取与乙醇诱导的 TER 减少、细胞高度降低和细胞间隙增加相关，这与之前的外部照射发现类似。低温提高了 TER，但尽管如此，冷却并没有改变颗粒的吸收。总之，培养微环境和假处理是体外上皮微粒摄取研究的关键考虑因素。 (C) 2009 Elsevier B.V. 保留所有权利。
Small intestinal microparticle uptake via a paracellular route is relevant to oral drug delivery and environmental pollution. In vitro investigation uses latex microparticle passage across a confluent Caco-2 cell epithelium. This paper examines the influence of culture conditions on transepithelial resistance (TER); cell dimensions from confocal microscopy; and number of particles below the epithelium. Variables investigated include level of initial TER; multiple TER measurements; involvement of medium; cell source; and pretreatment with ethanol or a range of temperatures. Data were collected after exposure to 2 mu m latex particles for 5-120 min: sham groups were exposed to pretreatment but not particles. The results highlight the importance of very precise control of the experimental environment: confirm the pattern of sequential-TER increase/decrease in groups exposed only to particles and show accompanying increases in cell dimensions. Greater particle uptake was associated with ethanol-induced decreased TER, decreased cell height and increased intercellular spaces, similar to previous findings for external irradiation. Low temperatures raised TER but, despite this, cooling did not alter particle uptake. In conclusion, culture microenvironment and sham treatment are crucial considerations in studies of epithelial microparticle uptake in vitro. (C) 2009 Elsevier B.V. All rights reserved.