Using reconfigurable microfluidics to study the role of HGF in autocrine and paracrine signaling of hepatocytes.
Using reconfigurable microfluidics to study the role of HGF in autocrine and paracrine signaling of hepatocytes.
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DOI：
10.1039/c5ib00105f
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发表时间：
2015-07
期刊：
影响因子：
--
通讯作者：
Revzin A
中科院分区：
文献类型：
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作者：
Patel D;Haque A;Gao Y;Revzin A
癌症、发育生物学和组织损伤提供了多个例子，其中邻近的细胞群通过旁分泌因子进行交流。在体内剖析这种细胞相互作用几乎是不可能的，在体外则相当具有挑战性。本研究的目的是利用可重构的微流体装置来研究体外原代肝细胞群之间的旁分泌信号交换。之前，我们证明位于含有胶原/HGF 点的蛋白点上的肝细胞表达上皮（肝）表型，并且还挽救了未接受直接 HGF 刺激的胶原点上的邻近肝细胞中的上皮表型。在此，我们设计了一种微流体装置，其具有由可伸缩（可重构）壁隔开的平行流体通道，并利用该装置来研究肝细胞组之间的相互作用，其中一些肝细胞受到 HGF 刺激，另一些则不受刺激。使用新型可重构微流体装置，我们证明在含有 HGF 的蛋白点上培养可以上调肝细胞中内源性 HGF 的产生，并且这些 HGF 分子扩散，导致受体细胞的表型增强。我们还表明，用 c-met 抑制剂 (SU11274) 选择性治疗受体肝细胞会减弱救援效果，这一点通过白蛋白和 HGF 表达的下调来衡量。我们的研究是第一个证明原代肝细胞中通过 HGF 进行旁分泌信号传导的研究之一。更广泛地说，这里描述的工具和方法可用于研究其他类型细胞中的旁分泌信号传导，并将与从癌症到免疫学的各个生物医学研究领域相关。
Cancer, developmental biology and tissue injury present multiple examples where groups of cells residing in close proximity communicate via paracrine factors. It is nearly impossible to dissect such cellular interactions in vivo and is quite challenging in vitro. The goal of this study is to utilize a reconfigurable microfluidic device in order to study paracrine signal exchange between groups of primary hepatocytes in vitro. Previously, we demonstrated that hepatocytes residing on protein spots containing collagen/HGF spots expressed epithelial (hepatic) phenotype and also rescued epithelial phenotype in neighboring hepatocytes on collagen spots that did not receive direct HGF stimulus. Herein, we designed a microfluidic device with parallel fluidic channels separated by retractable (reconfigurable) walls and employed this device to investigate interactions between groups of hepatocytes, some stimulated with HGF others not stimulated. Using novel reconfigurable microfluidic device, we demonstrate that cultivation on HGF-containing protein spots upregulates production of endogenous HGF in hepatocytes and that these HGF molecules diffuse over, causing phenotype enhancement in the recipient cells. We also show that selective treatment of the recipient hepatocytes with c-met inhibitor (SU11274) diminishes the rescue effect, as gauged by the down-regulation of albumin and HGF expression. Our study is one of the first to demonstrate paracrine signaling via HGF in primary hepatocytes. More broadly, tools and methods described here may be used to study paracrine signaling in other types of cells and will have relevance for various fields of biomedical research from cancer to immunology.