Magnetophoretic Cell Sorting and Analysis
Principal Investigator: Maciej Zborowski
Abstract: 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. PUBLIC HEALTH RELEVANCE: There is an intense interest in identifying cancer biomarkers for improved diagnosis and prognosis in the treatment of the disease. One particularly promising application is the detection, enumeration and analysis of circulating tumor cells (CTC's) in blood. There is evidence of abnormal paramagnetic contributions to the magnetic susceptibility of cells that have undergone malignant transformation coming from the laboratories collaborating on this grant application. We propose to systematically investigate this effect by using our state-of-the-art magnetic research facilities that include the magnetic cell separation laboratory and electron (spin) paramagnetic resonance (EPR) spectrometry facility available at the primary (Cleveland Clinic) and the subcontract (The Ohio State University) sites. The proposed research could have a profound impact on the magnetic separation methods in application to biomedical research and the treatment of cancer.
Funding Period: 1994-02-18 - 2016-01-31
more information: NIH RePORT
- Hemoglobin degradation in malaria-infected erythrocytes determined from live cell magnetophoresisLee R Moore
Department of Biomedical Engineering, Lerner Research Institute, The Cleveland Clinic Foundation, Cleveland, Ohio 44195, USA
FASEB J 20:747-9. 2006..50, in agreement with the published biochemical and crystallography data. Magnetophoretic analysis of live erythrocytes could become significant for antimalarial drug susceptibility and resistance determination...
- Quantification of non-specific binding of magnetic micro- and nanoparticles using cell tracking velocimetry: Implication for magnetic cell separation and detectionJ J Chalmers
William G Lowrie Department of Chemical and Biomolecular Engineering, The Ohio State University, 140 West 19th Avenue, Columbus, OH 43210, USA
Biotechnol Bioeng 105:1078-93. 2010....
- Sequential CD34 cell fractionation by magnetophoresis in a magnetic dipole flow sorterThomas Schneider
Department of Biomedical Engineering, Lerner Research Institute, Cleveland Clinic, 9500 Euclid Avenue, Cleveland, Ohio 44195, USA
Analyst 135:62-70. 2010..The fractional cell distributions showed good agreement with numerical simulations of the fractionation based on the cell magnetophoretic mobility distribution in the unsorted sample...
- Characterization of magnetic nanoparticles using programmed quadrupole magnetic field-flow fractionationP Stephen Williams
Department of Biomedical Engineering, Lerner Research Institute, Cleveland Clinic, 9500 Euclid Avenue, Cleveland, OH 44195, USA
Philos Trans A Math Phys Eng Sci 368:4419-37. 2010..The results are shown to be consistent and insensitive to conditions, although magnetite content appeared to be somewhat higher than expected...
- Quantification of both the presence, and oxidation state, of Mn in Bacillus atrophaeus spores and its imparting of magnetic susceptibility to the sporesJianxin Sun
William G Lowrie Department of Chemical and Biomolecular Engineering, The Ohio State University, Columbus, Ohio 43210, USA
Biotechnol Bioeng 108:1119-29. 2011..16 × 10(-11) g, which given the complexity of the two techniques, is a reasonable agreement. Finally, a discussion of potential applications of the magnetic properties of these spores is presented...
- Quantification of changes in oxygen release from red blood cells as a function of age based on magnetic susceptibility measurementsXiaoxia Jin
William G Lowrie Department of Chemical and Biomolecular Engineering, The Ohio State University, 140 W 19th Avenue, Columbus, OH 43210, USA
Analyst 136:2996-3003. 2011..The decrease in magnetophoretic mobility of the deoxyHb portion is explicable either by Hb's increased affinity for oxygen during storage, or else a loss of iron from the cells...
- Rare cell separation and analysis by magnetic sortingMaciej Zborowski
Department of Biomedical Engineering ND 20, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio 44195, United States
Anal Chem 83:8050-6. 2011....
- Iron transport in cancer cell culture suspensions measured by cell magnetophoresisXiaoxia Jin
Department of Biomedical Engineering ND20, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio 44195, United States
Anal Chem 84:4520-6. 2012....
- Emerging technologies for CTC detection based on depletion of normal cellsMaryam Lustberg
Internal Medicine, Division of Medical Oncology, The Ohio State University, Columbus, OH 43210, USA
Recent Results Cancer Res 195:97-110. 2012..In contrast to a positive selection methodology, this approach provides an unbiased characterization of these cells, including markers associated with epithelial mesenchymal transition...
- Multiparameter analysis, including EMT markers, on negatively enriched blood samples from patients with squamous cell carcinoma of the head and neckPriya Balasubramanian
William G Lowrie Department of Chemical and Biomolecular Engineering, The Ohio State University, Columbus, Ohio, United States of America
PLoS ONE 7:e42048. 2012....
- Erythrocyte enrichment in hematopoietic progenitor cell cultures based on magnetic susceptibility of the hemoglobinXiaoxia Jin
Department of Biomedical Engineering, Cleveland Clinic, Cleveland, Ohio, United States of America
PLoS ONE 7:e39491. 2012..The results demonstrate feasibility of label-free magnetic enrichment of erythrocyte fraction of CD34+ progenitor-derived cultures based on the presence of paramagnetic hemoglobin in the maturing erythrocytes...
- A quantitative determination of magnetic nanoparticle separation using on-off field operation of quadrupole magnetic field-flow fractionation (QMgFFF)Toru Orita
Division of Chemistry for Materials, Graduate School of Engineering, Mie University, Tsu, Mie, Japan
Anal Sci 29:761-4. 2013..This approach could be useful for detection of unwanted particulate contaminants, especially important in industrial and biomedical applications. ..
- Isolation and analysis of rare cells in the blood of cancer patients using a negative depletion methodologyYongqi Wu
William G Lowrie Department of Chemical and Biomolecular Engineering, The Ohio State University, Columbus, OH 43210, United States
Methods 64:169-82. 2013....
- Magnetic nanoparticle drug carriers and their study by quadrupole magnetic field-flow fractionationP Stephen Williams
Department of Biomedical Engineering, Lerner Research Institute, Cleveland Clinic, 9500 Euclid Avenue, Cleveland, Ohio 44195, USA
Mol Pharm 6:1290-306. 2009..Preliminary experiments involving a novel method of refining a magnetic nanoparticle drug carrier to achieve this result are described. QMgFFF is used to characterize the refined and unrefined material...
- Confocal images of circulating tumor cells obtained using a methodology and technology that removes normal cellsPriya Balasubramanian
William G Lowrie Department of Chemical and Biomolecular Engineering, The Ohio State University, 125 Koffolt Laboratories, 140 West 19th Avenue, Columbus, Ohio 43210, USA
Mol Pharm 6:1402-8. 2009..Future studies will involve the investigation if cancer stem cell and mesenchymal markers are present on these CTCs and correlations of patient outcome to the number and type of CTC present...
- Comparison of two immunomagnetic separation technologies to deplete T cells from human blood samplesOscar Lara
Department of Chemical and Biomolecular Engineering, The Ohio State University, 120 Koffolt Laboratories, 140 W 19th Avenue, Columbus, 43210, USA
Biotechnol Bioeng 94:66-80. 2006....
- Diagnosis of malaria by magnetic deposition microscopyPeter A Zimmerman
Case Western Reserve University, Center for Global Health and Diseases, Cleveland, Ohio 44106 7286, USA
Am J Trop Med Hyg 74:568-72. 2006..These results provide insight regarding new strategies for performing malaria blood smear microscopy...
- Continuous flow magnetic cell fractionation based on antigen expression levelThomas Schneider
Department of Biomedical Engineering, The Cleveland Clinic Foundation, 9500 Euclid Avenue, Cleveland, OH 44195, USA
J Biochem Biophys Methods 68:1-21. 2006....
- Binding affinities/avidities of antibody-antigen interactions: quantification and scale-up implicationsHuading Zhang
Department of Chemical and Biomolecular Engineering, The Ohio State University, 140 W 19th Ave, Columbus, Ohio 43210, USA
Biotechnol Bioeng 95:812-29. 2006....
- Blood progenitor cell separation from clinical leukapheresis product by magnetic nanoparticle binding and magnetophoresisYing Jing
Department of Biomedical Engineering, Lerner Research Institute, Cleveland Clinic, 9500 Euclid Ave, Cleveland, Ohio 44195, USA
Biotechnol Bioeng 96:1139-54. 2007..7-9.3) x 10(4) cells/s. The results also showed a highly regular dependence of the QMS performance on the flow conditions that agreed with the theoretical predictions based on the CD34+ cell magnetophoretic mobility...
- Application of immunomagnetic cell enrichment in combination with RT-PCR for the detection of rare circulating head and neck tumor cells in human peripheral bloodXiaodong Tong
Department of Chemical and Biomolecular Engineering, The Ohio State University, Columbus, Ohio 43210, USA
Cytometry B Clin Cytom 72:310-23. 2007..Using a single step immunomagnetic labeling, the final, optimized enrichment process produced a 57.6 +/- 30.3-fold (n = 6) enrichment of the rare cancer cells with a final cancer cell recovery of (77.8 +/- 6.6)%...
- Quantification of magnetic susceptibility in several strains of Bacillus spores: implications for separation and detectionKristie Melnik
Department of Chemical and Biomolecular Engineering, The Ohio State University, 140 West 19th Ave, Columbus, Ohio 43210, USA
Biotechnol Bioeng 98:186-92. 2007..These results indicate the potential to separate spores with intrinsic magnetic susceptibility directly out of water or air samples...
- Negative selection of hematopoietic progenitor cells by continuous magnetophoresisYing Jing
Department of Biomedical Engineering, Cleveland Clinic, 9500 Euclid Avenue, Cleveland, OH 44195, USA
Exp Hematol 35:662-72. 2007..To evaluate a negative selection technique for the hematopoietic progenitor cell enrichment from clinical leukapheresis product using continuous magnetophoresis...
- Complementary use of flow and sedimentation field-flow fractionation techniques for size characterizing biodegradable poly(lactic acid) nanospheresCatia Contado
Department of Chemistry, University of Ferrara, Via Luigi Borsari 46, I 44100 Ferrara, Italy
J Chromatogr A 1157:321-35. 2007..The two FFF techniques proved to be complementary and gave good, congruent and very useful information on the size distributions of the five poly(lactic acid) particle samples...
- A novel high throughput immunomagnetic cell sorting system for potential clinical scale depletion of T cells for allogeneic stem cell transplantationXiaodong Tong
Department of Chemical and Biomolecular Engineering, The Ohio State University, Columbus Ohio, USA
Exp Hematol 35:1613-22. 2007..0 log(10)), high level of recovery of hematopoietic stem cells (>90%), with a high throughput (>10(6) cells/second)...
- Utility of magnetic cell separation as a molecular sperm preparation techniqueTamer M Said
Department of Andrology and Reproductive Tissue Banking, Toronto Institute of Reproductive Medicine Repro Med, Toronto, Canada
J Androl 29:134-42. 2008..The higher sperm quality is represented by an increased oocyte penetration potential and cryosurvival rates. Thus, the selection of nonapoptotic spermatozoa by MACS should be considered to enhance ART success rates...
- Characterization of nonspecific crossover in split-flow thin channel fractionationP Stephen Williams
Department of Biomedical Engineering, Cleveland Clinic, 9500 Euclid Avenue, Cleveland, Ohio 44195, USA
Anal Chem 80:7105-15. 2008..It is concluded that SID may contribute to NSC, but that further experiments and mathematical modeling are necessary to more fully explore the phenomenon...
- Quantitative intracellular magnetic nanoparticle uptake measured by live cell magnetophoresisYing Jing
Department of Biomedical Engineering ND20, Cleveland Clinic, 9500 Euclid Ave, Cleveland, OH 44195, USA
FASEB J 22:4239-47. 2008..The results were consistent in all the cell types studied and with other reports. The cell magnetophoresis depends on the presence of high-spin iron species and is therefore expected to be directly related to the cell MRI contrast level...
- Optimization of an enrichment process for circulating tumor cells from the blood of head and neck cancer patients through depletion of normal cellsLiying Yang
Department of Chemical and Biomolecular Engineering, The Ohio State University, 125 Koffolt Laboratories, 140 West 19th Avenue, Columbus, Ohio 43210, USA
Biotechnol Bioeng 102:521-34. 2009..e., does the CTC have predominantly epithelial or mesenchymal characteristics?)...
- Development of multistage magnetic deposition microscopyPulak Nath
Department of Biomedical Engineering ND 20, Lerner Research Institute, Cleveland Clinic, 9500 Euclid Avenue, Cleveland, Ohio 44195, USA
Anal Chem 81:43-9. 2009..g., magnetic susceptibilities or magnetophoretic mobilities). The overall goal is to allow the screening of multiple disease conditions in a single platform...
- Differences in magnetically induced motion of diamagnetic, paramagnetic, and superparamagnetic microparticles detected by cell tracking velocimetryXiaoxia Jin
Department of Chemical and Biomolecular Engineering, The Ohio State University, Columbus, OH 43210, USA
Analyst 133:1767-75. 2008..The results demonstrate sensitivity of the CTV analysis to different magnetization mechanisms of the microparticles...
- Feasibility study of red blood cell debulking by magnetic field-flow fractionation with step-programmed flowLee R Moore
Department of Biomedical Engineering, Lerner Research Institute, Cleveland Clinic, 9500 Euclid Avenue, Cleveland, OH, 44195, USA
Anal Bioanal Chem 406:1661-70. 2014....