Isolation of Tumor Initiating Cells (TICs) using Contactless Dielectrophoresis

使用非接触式介电泳分离肿瘤起始细胞 (TIC)

• 批准号: 8547799
• 负责人: Scott D Cramer
• 金额: $ 18.73万
• 依托单位: VIRGINIA POLYTECHNIC INST AND ST UNIV
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-19 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8547799
• 关键词: Architecture; Behavior; Benign; Biological; Biological Assay; CD44 gene; Cancerous; Cell Separation; Cell Volumes; Cells; Clinical; Computer Simulation; Data; Development; Devices; Diagnostic; Effectiveness; Electrodes; Electrolyses; Environment; Exhibits; Fingerprint; Flow Cytometry; Frequencies; Future; Genotype; Goals; Human; In Vitro; Integrins; Label; Lead; Liquid substance; Methods; Microfluidics; Modeling; Monitor; Motion; Output; PC3 cell line; Pathogenesis; Patients; Performance; Physicians; Physics; Population; Procedures; Production; Property; Prostate; Relative (related person); Research Personnel; Role; Sampling; Solutions; Sorting - Cell Movement; Surface; System; Techniques; Technology; Testing; Therapeutic; Time; Tissues; Transplantation; Tumor Markers; aldehyde dehydrogenases; base; cancer cell; cancer stem cell; cancer therapy; cell killing; cell type; cost effectiveness; design; electric field; electrical property; human tissue; improved; in vivo; innovative technologies; killings; life time cost; neoplastic cell; novel; particle; physical property; prostate cancer cell; prostate cancer model; prototype; rapid technique; response; success; therapeutic target; tool; tumor; tumor growth; tumor progression; tumorigenic; voltage

DESCRIPTION (provided by applicant): Tumor initiating cells (TICs), also known as cancer stem cells, are a finite population of cancer cells that have the ability to transplant a new tumor from an existing one. They are also putatively responsible for the metastatic properties of tumors. Isolation of TICs is the first step towards understanding the role of these cells in the pathogenesis and progression of cancer and is critical towards the development of improved specific therapies for cancer. The TIC is the most relevant therapeutic target. However, current efforts in isolation and characterization of the TIC are hampered by the lack of suitable high throughput and rapid methods to isolate these cells. Contactless-dielectrophoresis (cDEP) is a new, non-invasive technique to detect and enrich rare cells suspended in a medium based on their physical and electrical properties, independent of their genotype. Dielectrophoresis (DEP) relies upon the motion of a particle due to its polarization induced by a non-uniform electric fiel. Conventional DEP microfluidic systems are susceptible to electrode fouling and require complicated fabrication procedures, because the electrodes are in direct contact with the sample, which limits their lifetime and cost effectiveness. cDEP alleviates these limitations by using external electrodes placed in secondary, highly conductive, channels that are separated from the cell sample by a thin insulating layer. Since the devices do not require complicated fabrication techniques, mass production is readily achievable and, therefore, this technique can be used by a large population of biologists and researchers for cell isolation. The application of DEP to separate target cells has been studied extensively in the last two decades, and the results indicate that rare cells can be separated from other cells in a cell mixture. Our central hypothesis is that cDEP is capable of discriminating between tumor imitating cells (TICs) and non-TICs. We will utilize a multifaceted approach involving experimentation and computational modeling to design new cDEP devices to assess and optimize the effectiveness of cDEP technology to physically separate and enrich tumor initiating cells from other cell populations found in a tumor. This multi-faceted approach will include 1) characterize the dielectrophoretic response of tumor initiating cells compared to typical tumor cells, 2) assess the performance of cDEP at isolating and selectively concentrating tumor initiating cells, 3) investigate possible subpopulation of TICs by sorting cells using cDEP microdevices, and 4) use primary human prostate tissue and repeat Aims 1, 2, and 3 with primary human prostate cells. Success in isolating TICs will lead to increased understanding of the most appropriate therapeutic strategies to ablate this cell population.

描述（由申请人提供）：肿瘤起始细胞（TIC），也称为癌症干细胞，是具有移植新肿瘤能力的有限癌细胞群 从现有的一个。它们也是肿瘤转移特性的主要原因。TIC的分离是理解这些细胞在癌症发病机制和进展中的作用的第一步，并且对于开发改进的癌症特异性疗法至关重要。TIC是最相关的治疗靶点。然而，目前在TIC的分离和表征方面的努力受到缺乏合适的高通量和快速分离这些细胞的方法的阻碍。非接触式介电电泳（cDEP）是一种新的非侵入性技术，用于检测和富集悬浮在培养基中的稀有细胞，基于它们的物理和电学性质，而不依赖于它们的基因型。介电电泳（DEP）依赖于粒子在非均匀电场诱导下的极化运动。传统的DEP微流体系统容易受到电极污染的影响，并且需要复杂的制造过程，因为电极与样品直接接触，这限制了它们的寿命和成本效益。cDEP通过使用放置在通过薄绝缘层与细胞样品分离的二级高导电通道中的外部电极来消除这些限制。由于该装置不需要复杂的制造技术，因此很容易实现大规模生产，因此，这种技术可以被大量生物学家和研究人员用于细胞分离。在过去的二十年中，DEP用于分离靶细胞的应用已经被广泛研究，并且结果表明可以将细胞混合物中的稀有细胞与其他细胞分离。我们的中心假设是，cDEP能够区分肿瘤模仿细胞（TIC）和非TIC。我们将利用涉及实验和计算建模的多方面方法来设计新的cDEP设备，以评估和优化cDEP技术的有效性，从而从肿瘤中发现的其他细胞群中物理分离和富集肿瘤起始细胞。这种多方面的方法将包括1）表征肿瘤起始细胞与典型肿瘤细胞相比的介电电泳响应，2）评估cDEP在分离和选择性浓缩肿瘤起始细胞方面的性能，3）通过使用cDEP微装置分选细胞来研究TIC的可能亚群，以及4）使用原代人前列腺组织并重复目的1，2，和3个用原代人前列腺细胞。分离TIC的成功将导致对消融该细胞群的最合适的治疗策略的理解增加。

项目成果

A feasibility study for enrichment of highly aggressive cancer subpopulations by their biophysical properties via dielectrophoresis enhanced with synergistic fluid flow.

一项可行性研究，可通过介电性流体增强其生物物理特性来富集高度侵略性的癌症亚群。

• DOI: 10.1002/elps.201600530
• 发表时间: 2017-06
• 期刊: Electrophoresis
• 影响因子: 2.9
• 作者: Douglas TA;Cemazar J;Balani N;Sweeney DC;Schmelz EM;Davalos RV
• 通讯作者: Davalos RV

Joule heating effects on particle immobilization in insulator-based dielectrophoretic devices.

焦耳热对基于绝缘体的介电泳装置中粒子固定的影响。

• DOI: 10.1002/elps.201300171
• 发表时间: 2014-02
• 期刊: ELECTROPHORESIS
• 影响因子: 2.9
• 作者: Gallo-Villanueva, Roberto C.;Sano, Michael B.;Lapizco-Encinas, Blanca H.;Davalos, Rafael V.
• 通讯作者: Davalos, Rafael V.

Improving cancer therapies by targeting the physical and chemical hallmarks of the tumor microenvironment.

• DOI: 10.1016/j.canlet.2015.12.019
• 发表时间: 2016-09-28
• 期刊: Cancer letters
• 影响因子: 9.7
• 作者: Ivey JW;Bonakdar M;Kanitkar A;Davalos RV;Verbridge SS
• 通讯作者: Verbridge SS

Modeling of Transmembrane Potential in Realistic Multicellular Structures before Electroporation.

• DOI: 10.1016/j.bpj.2016.10.005
• 发表时间: 2016-11
• 期刊: Biophysical journal
• 影响因子: 3.4
• 作者: T. Murovec;Daniel C. Sweeney;Eduardo L. Latouche;R. Davalos;C. Brosseau