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SGLT1 介导的 Caco-2 细胞转运对细胞库来源高度依赖。

SGLT1-Mediated Transport in Caco-2 Cells Is Highly Dependent on Cell Bank Origin.

机构信息

Department of Pharmacy, Faculty of Health and Medical Sciences, University of Copenhagen, Universitetsparken 2, Copenhagen DK-2100, Denmark; Department of Physics, Chemistry & Pharmacy, Faculty of Sciences, University of Southern Denmark, Campusvej 55, Odense DK-5230, Denmark.

Department of Pharmacy, Faculty of Health and Medical Sciences, University of Copenhagen, Universitetsparken 2, Copenhagen DK-2100, Denmark; Department of Physics, Chemistry & Pharmacy, Faculty of Sciences, University of Southern Denmark, Campusvej 55, Odense DK-5230, Denmark.

出版信息

J Pharm Sci. 2017 Sep;106(9):2664-2670. doi: 10.1016/j.xphs.2017.04.033. Epub 2017 Apr 26.

Abstract

The human colon adenocarcinoma (Caco-2) cell line is a well-established in vitro model for studying transport phenomena for prediction of intestinal nutrient and drug absorption. However, substances depending on transporters such predictions are complicated due to variable transporter expression and limited knowledge about transporter function during multiple cell passaging and cell thawings. In the case of sodium glucose transporter 1 (SGLT1), a key transporter of oral absorption of d-glucose, one reason for compromised prediction could be inadequate expression of SGLT1 in Caco-2 cells and thereby limited sensitivity in the determination of SGLT1-mediated permeability (P). Here, the objective is to characterize and compare SGLT1-mediated uptake in Caco-2 cells obtained from different cell banks. SGLT1-mediated uptake of the standard SGLT1 substrate, methyl-α-d-glucopyranoside, in Caco-2 cells was shown to be highly dependent on cell bank origin. The most robust and reliable SGLT1 functionality was identified in Caco-2 cells from Deutsche Sammlung für Mikroorganismen und Zellkulturen (DSMZ), whereas cells from the American Type Culture Collection and European Collection of Authenticated Cell Cultures have lower SGLT1 transport activity. Transepithelial P across Caco-2 cells from DSMZ showed that P likely accounts for approximately 97% of absorptive methyl-α-d-glucopyranoside P. In conclusion, Caco-2 cells from DSMZ provide a robust in vitro model for studying SGLT1-mediated uptake and transport-over multiple cell passages and independent cell stock thawings.

摘要

人结肠腺癌(Caco-2)细胞系是一种成熟的体外模型,可用于研究预测肠道营养物质和药物吸收的转运现象。然而,由于转运体表达的可变性以及在多次细胞传代和细胞解冻过程中对转运体功能的了解有限,依赖转运体的物质的预测变得复杂。在钠-葡萄糖共转运蛋白 1(SGLT1)的情况下,SGLT1 是口服吸收 D-葡萄糖的关键转运体,预测能力受损的原因之一可能是 Caco-2 细胞中 SGLT1 的表达不足,从而限制了 SGLT1 介导的通透性(P)的测定的灵敏度。在这里,我们的目的是对不同细胞库获得的 Caco-2 细胞中 SGLT1 介导的摄取进行特征描述和比较。结果表明,SGLT1 介导的标准 SGLT1 底物甲基-α-D-吡喃葡萄糖苷在 Caco-2 细胞中的摄取高度依赖于细胞库的来源。在 Deutsche Sammlung für Mikroorganismen und Zellkulturen(DSMZ)中获得的 Caco-2 细胞中,SGLT1 功能最稳健和可靠,而源自美国模式培养物集存库和欧洲细胞培养物保藏中心的细胞的 SGLT1 转运活性较低。来自 DSMZ 的 Caco-2 细胞的跨上皮 P 表明,P 可能约占吸收性甲基-α-D-吡喃葡萄糖苷 P 的 97%。总之,DSMZ 的 Caco-2 细胞提供了一种稳健的体外模型,可用于研究 SGLT1 介导的摄取和转运,可在多个细胞传代和独立细胞库解冻中进行。

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