A novel microfluidic co-culture system for investigation of bacterial cancer targeting
A novel microfluidic co-culture system for investigation of bacterial cancer targeting
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DOI：
10.1039/c3lc50163a
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发表时间：
2013-01-01
期刊：
影响因子：
6.1
通讯作者：
Shin, Jennifer H.
中科院分区：
文献类型：
--
作者：
Hong, Jung Woo;Song, Sukhyun;Shin, Jennifer H.
虽然以前已经证明并建议在动物模型中靶向细菌癌症作为一种可能的治疗工具，但在临床应用之前，需要彻底了解负责癌症特异性的机制。为了可视化细菌对癌细胞的偏好超过正常细胞并阐明癌症靶向机制，已经开发了一种简单的微流体平台用于体外研究。该平台允许在隔离室中的独立培养环境中同时培养多种细胞类型，并在每个细胞培养室和中央通道之间的胶原填充通道上产生稳定的化学梯度。所建立的化学梯度诱导细菌向特定细胞类型的趋化性优先迁移以进行定量分析。作为示范，我们在双室装置上测试了不同的细菌行为，其中我们基于在胶原填充通道的两个出口处表达绿色荧光蛋白（GFP）的细菌的荧光强度的差异来量化细菌偏好。使用开发的平台分析鼠伤寒沙门氏菌对正常肝细胞和癌肝细胞的趋化行为，发现对癌肝细胞有明显的偏好。我们还证明了甲胎蛋白（AFP）是沙门氏菌的关键化学引诱剂之一。鼠伤寒沙门氏菌治疗肝癌
Although bacterial cancer targeting in animal models has been previously demonstrated and suggested as a possible therapeutic tool, a thorough understanding of the mechanisms responsible for cancer specificity would be required prior to clinical applications. To visualize bacterial preference for cancer cells over normal cells and to elucidate the cancer-targeting mechanism, a simple microfluidic platform has been developed for in vitro studies. This platform allows simultaneous cultures of multiple cell types in independent culture environments in isolated chambers, and creates a stable chemical gradient across a collagen-filled passage between each of these cell culture chambers and the central channel. The established chemical gradient induces chemotactic preferential migration of bacteria toward a particular cell type for quantitative analysis. As a demonstration, we tested differential bacterial behavior on a two-chamber device where we quantified bacterial preference based on the difference in fluorescence intensities of green fluorescence protein (GFP)-expressing bacteria at two exits of the collagen-filled passages. Analysis of the chemotactic behavior of Salmonella typhimurium toward normal versus cancer hepatocytes using the developed platform revealed an apparent preference for cancer hepatocytes. We also demonstrate that alpha-fetoprotein (AFP) is one of the key chemo-attractants for S. typhimurium in targeting liver cancer.