A multi-chamber microfluidic intestinal barrier model using Caco-2 cells for drug transport studies.
A multi-chamber microfluidic intestinal barrier model using Caco-2 cells for drug transport studies.
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DOI：
10.1371/journal.pone.0197101
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发表时间：
2018
期刊：
影响因子：
3.7
通讯作者：
Andresen TL
中科院分区：
文献类型：
--
作者：
Tan HY;Trier S;Rahbek UL;Dufva M;Kutter JP;Andresen TL
本文介绍了一种使用硫醇-烯“点击化学”的多层多室微芯片系统的设计和制造，旨在研究药物在组织屏障模型中的传输。该制造工艺使用不同的硫醇-烯聚合物混合物实现了多层微流控芯片的快速成型，其中用于细胞单层生长的多孔特氟龙膜通过掩蔽的夹层硫醇-烯流体层结合在一起。用于跨上皮电阻(TEER)测量的电极使用低熔点焊丝与铂丝相结合，实现了对八个腔室的屏障完整性的并行实时监控。此外，半透明的多孔特氟龙膜能够对细胞单分子层进行光学监测。该装置是用Caco-2肠道模型开发和测试的，并与传统的Transwell系统进行了比较。通过紧密连接和粘液蛋白的原位免疫细胞化学、P-糖蛋白1(P-gp)介导的罗丹明123外排和刷状缘氨基肽酶活性检测细胞单层分化。通过甘露醇、右旋糖苷和胰岛素的渗透性研究，或与吸收促进剂十四烷基麦芽糖苷(TDM)联合使用，评价单层的紧密性和药物释放研究的相关性。基于硫醇-烯的微芯片材料和电极与细胞生长高度相容。事实上，在该设备中培养的Caco-2细胞在细胞培养的9-10天内显示出分化、粘液产生、定向运输和氨基肽酶活性，表明屏障形成的速度比传统Transwell模型更快。细胞单层表现出高的TEER和对亲水性化合物的致密性，而联合使用吸收促进剂引起TEER降低和通透性增加，与Transwell培养相似。提出的细胞屏障微设备构成了相关的组织屏障模型，使药物和化学品的传输研究能够在八个平行的腔室中进行实时光学和功能监测，从而提高了与以前报道的微设备相比的吞吐量。
This paper presents the design and fabrication of a multi-layer and multi-chamber microchip system using thiol-ene ‘click chemistry’ aimed for drug transport studies across tissue barrier models. The fabrication process enables rapid prototyping of multi-layer microfluidic chips using different thiol-ene polymer mixtures, where porous Teflon membranes for cell monolayer growth were incorporated by masked sandwiching thiol-ene-based fluid layers. Electrodes for trans-epithelial electrical resistance (TEER) measurements were incorporated using low-melting soldering wires in combination with platinum wires, enabling parallel real-time monitoring of barrier integrity for the eight chambers. Additionally, the translucent porous Teflon membrane enabled optical monitoring of cell monolayers. The device was developed and tested with the Caco-2 intestinal model, and compared to the conventional Transwell system. Cell monolayer differentiation was assessed via in situ immunocytochemistry of tight junction and mucus proteins, P-glycoprotein 1 (P-gp) mediated efflux of Rhodamine 123, and brush border aminopeptidase activity. Monolayer tightness and relevance for drug delivery research was evaluated through permeability studies of mannitol, dextran and insulin, alone or in combination with the absorption enhancer tetradecylmaltoside (TDM). The thiol-ene-based microchip material and electrodes were highly compatible with cell growth. In fact, Caco-2 cells cultured in the device displayed differentiation, mucus production, directional transport and aminopeptidase activity within 9–10 days of cell culture, indicating robust barrier formation at a faster rate than in conventional Transwell models. The cell monolayer displayed high TEER and tightness towards hydrophilic compounds, whereas co-administration of an absorption enhancer elicited TEER-decrease and increased permeability similar to the Transwell cultures. The presented cell barrier microdevice constitutes a relevant tissue barrier model, enabling transport studies of drugs and chemicals under real-time optical and functional monitoring in eight parallel chambers, thereby increasing the throughput compared to previously reported microdevices.
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影响因子：
3.7
作者：
Basson, MD;Turowski, G;Emenaker, NJ
通讯作者：
Emenaker, NJ
影响因子：
2.8
作者：
Chi, Meiying;Yi, Banya;Park, Sungsu
通讯作者：
Park, Sungsu
影响因子：
--
作者：
Carlborg, Carl Fredrik;Vastesson, Alexander;Haraldsson, Tommy
通讯作者：
Haraldsson, Tommy
影响因子：
3.9
作者：
Chumbimuni-Torres KY;Coronado RE;Mfuh AM;Castro-Guerrero C;Silva MF;Negrete GR;Bizios R;Garcia CD
通讯作者：
Garcia CD
影响因子：
4.1
作者：
HOWELL, S;KENNY, AJ;TURNER, AJ
通讯作者：
TURNER, AJ