An open microfluidic device to study chemotaxis of eukaryotic cells

用于研究真核细胞趋化性的开放式微流体装置

• 批准号: 8330999
• 负责人: Dawit Jowhar
• 金额: $ 2.48万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2013-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8330999
• 关键词: Academic Medical Centers; Actins; Address; Amoeba genus; Arthritis; Behavior; Biological Assay; Biological Models; Biosensor; Cell Polarity; Cells; Chemicals; Chemotactic Factors; Chemotaxis; Cytoskeletal Proteins; Development; Devices; Dictyostelium discoideum; Disease; Embryonic Development; Equipment; Eukaryotic Cell; Glass; Goals; Human Resources; Imagery; Immobilized Cells; Immune system; Inflammation; Knowledge; Laboratories; Lead; Learning; Literature; Location; Measures; Microfluidic Microchips; Microfluidics; Mission; Movement; National Institute of General Medical Sciences; Neoplasm Metastasis; Organ; Outcome; Output; Pathway interactions; Play; Primary Neoplasm; Process; Public Health; Qualifying; Race; Relative (related person); Research; Research Project Grants; Research Proposals; Research Technics; Role; Signal Transduction; Signaling Molecule; Signaling Protein; Speed; Techniques; Training; Work; cancer cell; cell motility; flexibility; improved; inhibitor/antagonist; migration; neutrophil; novel; pathogen; polarized cell; polydimethylsiloxane; polymerization; prevent; programs; research study; response; social; theories; therapy design; tissue repair; tumor

DESCRIPTION (provided by applicant): The proposed research project seeks to investigate the mechanisms by which cells polarize and help further our understanding of the role of polarization in directed cell migration. These studies will use the social amoeba Dictyostelium discoideum and will be conducted using a newly developed open microfluidic device made out of Polydimethylsiloxane. This device has multiple, micron-sized channels which can be used to race morphologically distinct cells against each other and the channels are also capable of confining cells for polarity studies. By learning more about D. discoideum cells, these findings can be correlated with the chemotaxis processes controlling other eukaryotic cells such as neutrophils and also elucidate the pathways that regulate cancer metastasis. In order to achieve these goals, three specific aims have been set. The first specific aim focuses on the further development of an open microfluidic devices that can be used for chemotaxis assays, and allows the visualization of a stable, passive chemical gradient. The second specific aim targets the role and mechanisms of polarity in chemotaxing cells by comparing the migration of polarized and unpolarized D. Discoideum cells towards their respective chemoattractants. Cells expressing fluorescent cytoskeletal and signaling proteins will be exposed to chemoattractant at one end of a microfluidic channel, after which the gradient will be reversed and the redistribution of these fluorescent molecules will be analyzed as the cells de-polarize and re-polarize. The third specific aim focuses on the gradient sensing mechanism in eukaryotic cells. The concentration of the gradient (input) will be compared to the migration speeds and signaling protein localizations (output). In line with the mission of the National Institute of General Medical Sciences (NIGMS), this research proposal seeks to understand the chemotaxis processes of eukaryotic cells. The findings from these experiments will help elucidate the basic mechanisms by which cells sense chemical gradients and migrate and further our understanding of how cancer cells metastasize. The proposed research program will require the P.I. to apply theory learned in both the classroom and from scientific literature. Appropriate research techniques will be picked up in the laboratory and the experiments will be performed under the guidance of the research sponsor. Training and implementation on the use of appropriate equipment and techniques will be facilitated by qualified personnel in the Janetopoulos laboratory and the staff at Vanderbilt University medical center.

描述（由申请人提供）：拟议的研究项目旨在研究细胞极化的机制，并帮助我们进一步了解极化在定向细胞迁移中的作用。这些研究将使用社会阿米巴盘基网柄菌，并使用新开发的由聚二甲基硅氧烷制成的开放式微流体装置进行。该装置具有多个微米大小的通道，可用于使形态不同的细胞相互竞争，并且这些通道还能够限制细胞进行极性研究。通过更多地了解盘状 D. discoideum 细胞，这些发现可以与控制其他真核细胞（如中性粒细胞）的趋化过程相关联，并阐明调节癌症转移的途径。为了实现这些目标，制定了三个具体目标。第一个具体目标侧重于进一步开发可用于趋化性测定的开放式微流体装置，并允许稳定、被动化学梯度的可视化。第二个具体目标是通过比较极化和非极化的盘状圆盘藻细胞向各自趋化剂的迁移来研究极性在趋化细胞中的作用和机制。表达荧光细胞骨架和信号蛋白的细胞将在微流体通道的一端暴露于化学引诱剂，之后梯度将反转，并且随着细胞去极化和重新极化，将分析这些荧光分子的重新分布。第三个具体目标集中于真核细胞中的梯度传感机制。将梯度浓度（输入）与迁移速度和信号蛋白定位（输出）进行比较。根据国家普通医学科学研究所 (NIGMS) 的使命，该研究计划旨在了解真核细胞的趋化过程。这些实验的结果将有助于阐明细胞感知化学梯度和迁移的基本机制，并进一步了解癌细胞如何转移。拟议的研究计划将需要 P.I.应用在课堂和科学文献中学到的理论。将在实验室中采用适当的研究技术，并在研究资助者的指导下进行实验。 Janetopoulos 实验室的合格人员和范德比尔特大学医学中心的工作人员将促进有关使用适当设备和技术的培训和实施。