Magnetophoretic Cell Sorting and Analysis

磁泳细胞分选和分析

• 批准号: 8795167
• 负责人: Maciej Zborowski
• 金额: $ 35.75万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-02-18 至 2017-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8795167
• 关键词: Appearance; Applications Grants; Biological Markers; Biomedical Research; Blood; Blood Cells; Cancer cell line; Cell Separation; Cells; Clinic; Collaborations; Complement; Computer Assisted; Computer software; Culture Media; Data; Detection; Diagnosis; Disease; Electrolytes; Electron Spin Resonance Spectroscopy; Electrons; Erythrocytes; Exhibits; Ferritin; Field Flow Fractionation; Funding; Health; Hela Cells; Heme; Human; Image; Immunophenotyping; Iron; Iron Compounds; Label; Laboratories; Life; Link; Liquid substance; Magnetic Resonance Imaging; Magnetism; Malignant - descriptor; Malignant Neoplasms; Manganese; Mass Spectrum Analysis; Measures; Methods; Modeling; Molecular; Morphologic artifacts; Motion; Neoplasm Circulating Cells; Normal Cell; Ohio; Oncologist; Organic Iron Compounds; Particulate; Physiological; Predisposition; Primary Neoplasm; Property; Proteins; Protocols documentation; Reactive Oxygen Species; Research; Resolution; Rest; Site; Solid Neoplasm; Solutions; Spectrometry; Spectrum Analysis; Structure; Suspension substance; Suspensions; Technology; Testing; Tissues; Transferrin Receptor; Transport Process; Tumor Cell Line; Universities; Velocimetries; Whole Blood; absorption; analytical method; base; cancer cell; cancer therapy; cell analyzer; cell transformation; cytochrome c; energy density; improved; instrumentation; interest; magnetic beads; magnetic cell separation; magnetic field; magnetic sorter; neoplastic cell; outcome forecast; receptor expression; research facility; uptake

DESCRIPTION (provided by applicant): This is a competing renewal application that builds on the instrumentation and expertise developed in the course of the previous funding periods. We propose to investigate the feasibility of extending the current magnetic cell separation methods to applications relying on the cell's own, natural (intrinsic) magnetization. The application rests on the hypothesis that there is a link between malignant cell transformation and an increase in the cell magnetic susceptibility. The supporting evidence comes from a large body of data from other studies on the magnetic properties of tissues, in particular, from electron (spin) paramagnetic resonance, EPR, and studies of solid tumors. Additional supporting evidence comes from our own data on the magnetic field-induced motion (magnetophoresis) in physiologic electrolyte solutions of cancer cell lines (HeLa and Hep 3B), that demonstrate statistically greater velocity compared to controls (oxygenated red blood cells). We have shown an additional increase in the cell magnetophoretic mobility following the addition of soluble iron compounds to the culture media, suggesting an effect of intracellular iron uptake on the cell magnetization. The proposed effort is divided between: Specific Aims: SA1: To investigate the molecular mechanism for the observed increase in magnetophoretic mobility (MM) in selected cancer cell lines as compared to matched normal cells. In collaboration with experts in cell tracking velocimetry (Dr. Chalmers) and EPR spectrometry (Dr. Kuppusamy) at the subcontract site (The Ohio State University), the effect of iron in the medium on the cell magnetophoretic mobility and the increased intracellular paramagnetic content will be investigated on cancer cell lines. SA2: To improve the sensitivity and resolution of the cell magnetophoretic mobility analyzer, the Cell Tracking Velocimetry (CTV). The increase of magnetophoretic mobility resolution by a factor up to 10 fold will be accomplished by increasing the magnetic energy density and field gradient of the apparatus, by improving the imaging capability of the cell tracking hardware and software, and by including corrections for fluid dynamics artifacts in collaboration with an expert in field-flow fractionation (Dr. Williams). SA3: To measure the magnetic susceptibility of selected primary tumors at a single cell level against a baseline of normal blood cells. Primary tumor cell lines will be tested against normal controls for differences in the magnetophoretic mobility, in collaboration with an expert oncologist (Dr. Borden). SA4: To test the feasibility of the label-less, magnetic cancer cell separation on model blood cell suspensions. The proposed strategy is based on the soluble - rather than particulate - iron transport processes, and may therefore be more finely tuned to the pathobiology of the cancer cell than is practiced today. It could have a profound impact on how the magnetic cell separation is practiced and utilized and could complement and extend the applications of the already existing magnetic cell separation methods.

描述（由申请人提供）：这是一个竞争性的续期申请，建立在以前资助期间开发的仪器和专业知识的基础上。我们建议调查的可行性，目前的磁性细胞分离方法的应用程序依赖于细胞自身的，自然的（内在的）磁化。该应用依赖于恶性细胞转化和细胞磁化率增加之间存在联系的假设。支持证据来自于其他组织磁特性研究的大量数据，特别是电子（自旋）顺磁共振、EPR和实体瘤研究。其他支持证据来自我们自己的数据，磁场诱导的运动（磁泳）在生理电解质溶液中的癌细胞系（HeLa和Hep 3B），这表明统计学上更大的速度相比，控制（含氧红细胞）。我们已经表明，在培养基中添加可溶性铁化合物后，细胞磁泳迁移率进一步增加，这表明细胞内铁吸收对细胞磁化的影响。具体目标：SA 1：研究与匹配的正常细胞相比，在选定的癌细胞系中观察到的磁泳迁移率（MM）增加的分子机制。与研究中心（俄亥俄州州立大学）的细胞跟踪测速法（查尔默斯博士）和EPR光谱法（Kuppusamy博士）专家合作，将在癌细胞系上研究培养基中的铁对细胞磁泳迁移率和细胞内顺磁性含量增加的影响。SA 2：为了提高细胞磁泳迁移率分析仪的灵敏度和分辨率，细胞跟踪测速（CTV）。通过增加装置的磁能密度和磁场梯度，通过改善细胞跟踪硬件和软件的成像能力，以及通过与场流分级专家（威廉姆斯博士）合作对流体动力学伪影进行校正，可以将磁泳迁移率分辨率提高10倍。SA 3：在单细胞水平上测量选定原发性肿瘤相对于正常血细胞基线的磁化率。将与肿瘤专家（Borden博士）合作，针对磁泳迁移率的差异，对原发性肿瘤细胞系与正常对照进行检测。SA 4：在模型血细胞悬液上测试无标记磁性癌细胞分离的可行性。所提出的策略是基于可溶性-而不是颗粒-铁运输过程，因此可能比今天更精细地调整到癌细胞的病理生物学。它可能对磁性细胞分离的实践和利用产生深远的影响，并可能补充和扩展现有磁性细胞分离方法的应用。

项目成果

Optimization of an enrichment process for circulating tumor cells from the blood of head and neck cancer patients through depletion of normal cells.

• DOI: 10.1002/bit.22066
• 发表时间: 2009-02-01
• 期刊: BIOTECHNOLOGY AND BIOENGINEERING
• 影响因子: 3.8
• 作者: Yang, Liying;Lang, James C.;Balasubramanian, Priya;Jatana, Kris R.;Schuller, David;Agrawal, Amit;Zborowski, Maciej;Chalmers, Jeffrey J.
• 通讯作者: Chalmers, Jeffrey J.

Magnetic flow sorting using a model system of human lymphocytes and a colloidal magnetic label.

使用人类淋巴细胞模型系统和胶体磁性标记进行磁流分选。

• DOI: 10.1097/00002480-199609000-00071
• 发表时间: 1996
• 期刊: ASAIO journal (American Society for Artificial Internal Organs : 1992)
• 作者: Zborowski,M;Moore,LR;Reddy,S;Chen,GH;Sun,L;Chalmers,JJ
• 通讯作者: Chalmers,JJ

Rare cell separation and analysis by magnetic sorting.

• DOI: 10.1021/ac200550d
• 发表时间: 2011-11-01
• 期刊: ANALYTICAL CHEMISTRY
• 影响因子: 7.4
• 作者: Zborowski, Maciej;Chamers, Jeffrey J.
• 通讯作者: Chamers, Jeffrey J.

Quantification of non-specific binding of magnetic micro- and nanoparticles using cell tracking velocimetry: Implication for magnetic cell separation and detection.

• DOI: 10.1002/bit.22635
• 发表时间: 2010-04-15
• 期刊: BIOTECHNOLOGY AND BIOENGINEERING
• 影响因子: 3.8
• 作者: Chalmers, J. J.;Xiong, Y.;Jin, X.;Shao, M.;Tong, X.;Farag, S.;Zborowski, M.
• 通讯作者: Zborowski, M.

Heterogeneous atypical cell populations are present in blood of metastatic breast cancer patients.

• DOI: 10.1186/bcr3622
• 发表时间: 2014-03-06
• 期刊: Breast cancer research : BCR
• 作者: Lustberg MB;Balasubramanian P;Miller B;Garcia-Villa A;Deighan C;Wu Y;Carothers S;Berger M;Ramaswamy B;Macrae ER;Wesolowski R;Layman RM;Mrozek E;Pan X;Summers TA;Shapiro CL;Chalmers JJ
• 通讯作者: Chalmers JJ