A microfluidic cell culture device (μFCCD) to culture epithelial cells with physiological and morphological properties that mimic those of the human intestine
A microfluidic cell culture device (μFCCD) to culture epithelial cells with physiological and morphological properties that mimic those of the human intestine
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DOI：
10.1007/s10544-015-9966-5
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发表时间：
2015-06-01
影响因子：
2.8
通讯作者：
Park, Sungsu
中科院分区：
文献类型：
--
作者：
Chi, Meiying;Yi, Banya;Park, Sungsu
人肠的生理学和形态学特性不能在常规培养装置如孔板和皮氏培养皿中准确地模拟，其中肠上皮细胞形成单层，细胞之间接触松散。在这里，我们报告了一种新的微流控细胞培养装置（亩FCCD），可用于培养细胞作为人类肠道模型。该装置使肠上皮细胞（Caco-2）能够在两个聚二甲基硅氧烷（PDMS）层之间涂覆有纤连蛋白的多孔膜上三维生长。在3天内，Caco-2细胞在mu FCCD中培养形成绒毛和隐窝样结构，具有小的细胞间空间，而单个细胞通过表达紧密连接蛋白occludin彼此紧密连接，并被分泌的粘蛋白MUC-2覆盖。在mu FCCD中培养3天的Caco-2细胞对细菌攻击的敏感性低于在transwell平板中培养21天的Caco-2细胞。μ FCCD培养的Caco-2细胞也显示出生理相关的吸收和细胞旁转运特性。这些结果表明，我们的肠道模型比传统的transwell培养模型更准确地模拟了体内肠道的形态和生理特性。
Physiological and morphological properties of the human intestine cannot be accurately mimicked in conventional culture devices such as well plates and petri dishes where intestinal epithelial cells form a monolayer with loose contacts among cells. Here, we report a novel microfluidic cell culture device (mu FCCD) that can be used to culture cells as a human intestinal model. This device enables intestinal epithelial cells (Caco-2) to grow three-dimensionally on a porous membrane coated with fibronectin between two polydimethylsiloxane (PDMS) layers. Within 3 days, Caco-2 cells cultured in the mu FCCD formed villi-and crypt-like structures with small intercellular spaces, while individual cells were tightly connected to one another through the expression of the tight junction protein occludin, and were covered with a secreted mucin, MUC-2. Caco-2 cells cultured in the mu FCCD for 3 days were less susceptible to bacterial attack than those cultured in transwell plates for 21 days. mu FCCD-cultured Caco-2 cells also displayed physiologically relevant absorption and paracellular transport properties. These results suggest that our intestinal model more accurately mimics the morphological and physiological properties of the intestine in vivo than the conventional transwell culture model.