Xiuwen Wu, Ru-Feng Wang, Ming Yuan, W. Xu, Xiu-wei Yang
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引用次数: 14
摘要
Caco-2细胞单层模型广泛应用于药物吸收研究。Dulbecco's Modified Eagle's Medium (DMEM)和Minimum Essential Medium (MEM)在该模型的开发中被交替使用,尽管它们的成分不同,可能会影响Caco-2细胞单层的分化和连接形成。本文比较了两种培养基中培养的Caco-2细胞单层,以奠定该模型的标准化基础。通过形态学、经上皮电阻(TEER)、碱性磷酸酶(AKPase)活性和转运实验,比较评价了这两种单层膜的可靠性和稳定性。虽然结果显示,两种单分子膜的顶端侧均存在特征微绒毛,但DMEM培养的单分子膜TEER的动态变化比MEM培养的更稳定,且前者的AKPase活性强于后者。此外,通常用于评估该模型的关键指标阿替洛尔(atenolol)在MEM培养的单层中的数量明显多于DMEM培养的单层。因此,DMEM培养的Caco-2单层膜比MEM培养的Caco-2单层膜更可靠、更稳定,因此DMEM培养的Caco-2单层膜更适合体外药物吸收研究。
Dulbecco's modified eagle's medium and minimum essential medium--which one is more preferred for establishment of Caco-2 cell monolayer model used in evaluation of drug absorption?
The Caco-2 cell monolayer model is widely used in drug absorption studies. Dulbecco's Modified Eagle's Medium (DMEM) and Minimum Essential Medium (MEM) have been used alternatively in the development of this model, although they are different in composition which may affect the differentiation and junction formation of the Caco-2 cell monolayer. Two Caco-2 cell monolayers cultured in both media were compared herein in order to underlay the standardization of this model. These two monolayers were comparatively evaluated regarding reliability and stability by morphology, transepithelial electrical resistance (TEER), alkaline phosphatase (AKPase) activity and transport experiments. Although the results showed that characteristic microvilli were present at the apical side of both monolayers, the dynamic change of TEER of the monolayer cultured in DMEM was more stable than that cultured in MEM, and AKPase activity of the former was stronger than that of the latter. Furthermore, the quantity of atenolol, a key indicator usually used for assessment of this model, across the monolayer cultured in MEM was significantly more than that cultured in DMEM. Therefore, the Caco-2 monolayer cultured in DMEM was more reliable and stable than that cultured in MEM, and thus the former was preferred for drug absorption investigation in vitro.