Separation of cells based on their function or surface properties is a basis of many techniques used in cell biology, cancer therapy and in biotechnology. While immunomagnetic cell separation has been utilized in all these areas, it has been hampered by the relatively large size of the paramagnetic label as compared to the target cell and by the relatively crude devices used for separation. This study will have two primary goals: the development of a novel magnetic cell label, and the analysis and design of a continuous, magnetic cell sorter using this label. This new magnetic label consists of ferritin combined to monoclonal antibodies against a particular surface antigen on the target cell. Ferritin is an iron storage protein in mammals. The properties of ferritin labels will be characterized using a number of techniques including: nuclear magnetic resonance spectroscopy, atomic absorption spectroscopy, and transmission electron microscopy. A unique sandwich labeling technique combining fluorescent and magnetic label will be used in combination with flow cytometry to characterize the cell-label binding. Human peripheral lymphocytes and human breast carcinoma derived cells will be used in this study, in combination with monoclonal antibodies against the specific markers: CD3, CD4, CD8, CD16, CD34 and Ber-EP4. These markers identify cells important for the bone marrow transplantation therapy: T cell, helper/inducer, cytotoxic/suppressor, natural killer, stem and breast carcinoma cells, respectively. Due to the complexity involved, the analysis and design of a continuous, magnetic cell sorter will require the use of 3-Dimensional Particle Image Velocimetry (3-D PIV). 3-D PIV, developed at the Ohio State University, the proposed subcontract site of this study, will allow the determination of 3-D velocity vectors of labeled and unlabeled cells in high magnetic fields. The experimental analysis, combined with the relevant equations of motion, will provide information on the system at a micron scale. With this information an optimized cell sorter will be developed. In addition, it will be possible to design more advanced sorters, such as multistage system, in which very high purity or very high selectivity can be obtained in a relatively short time. The ultimate goal is to develop a continuous, magnetic cell sorter for large-scale separation for bone marrow transplantation therapies. Such a sorter will be applicable to rapid, large-scale T cell depletion, cancer cell purging and hematopoietic stem cell enrichment.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
1R01CA062349-01
Application #
2103509
Study Section
Special Emphasis Panel (ZRG7-SSS-3 (08))
Project Start
1994-02-18
Project End
1997-01-31
Budget Start
1994-02-18
Budget End
1995-01-31
Support Year
1
Fiscal Year
1994
Total Cost
Indirect Cost
Name
Cleveland Clinic Lerner
Department
Type
DUNS #
017730458
City
Cleveland
State
OH
Country
United States
Zip Code
44195
Chalmers, J J; Jin, X; Palmer, A F et al. (2017) Femtogram Resolution of Iron Content on a Per Cell Basis: Ex Vivo Storage of Human Red Blood Cells Leads to Loss of Hemoglobin. Anal Chem 89:3702-3709
Mahajan, Kalpesh D; Nabar, Gauri M; Xue, Wei et al. (2017) Mechanotransduction Effects on Endothelial Cell Proliferation via CD31 and VEGFR2: Implications for Immunomagnetic Separation. Biotechnol J 12:
Moore, Lee R; Williams, P Stephen; Chalmers, Jeffrey J et al. (2017) Tessellated permanent magnet circuits for flow-through, open gradient separations of weakly magnetic materials. J Magn Magn Mater 427:325-330
Wu, Yongqi; Park, Kyoung-Joo Jenny; Deighan, Clayton et al. (2016) Multiparameter Evaluation of the Heterogeneity of Circulating Tumor Cells Using Integrated RNA In Situ Hybridization and Immunocytochemical Analysis. Front Oncol 6:234
Swaminathan, Ganesh; Sivaraman, Balakrishnan; Moore, Lee et al. (2016) Magnetically Responsive Bone Marrow Mesenchymal Stem Cell-Derived Smooth Muscle Cells Maintain Their Benefits to Augmenting Elastic Matrix Neoassembly. Tissue Eng Part C Methods 22:301-11
Joshi, Powrnima; Kooshki, Mitra; Aldrich, Wayne et al. (2016) Expression of natural killer cell regulatory microRNA by uveal melanoma cancer stem cells. Clin Exp Metastasis 33:829-838
Sumari, Deborah; Grimberg, Brian T; Blankenship, D'Arbra et al. (2016) Application of magnetic cytosmear for the estimation of Plasmodium falciparum gametocyte density and detection of asexual stages in asymptomatic children. Malar J 15:113
Buck, Amy; Moore, Lee R; Lane, Christopher D et al. (2015) Magnetic separation of algae genetically modified for increased intracellular iron uptake. J Magn Magn Mater 380:201-204
Joshi, Powrnima; Williams, P Stephen; Moore, Lee R et al. (2015) Circular Halbach array for fast magnetic separation of hyaluronan-expressing tissue progenitors. Anal Chem 87:9908-15
Lustberg, Maryam B; Balasubramanian, Priya; Miller, Brandon et al. (2014) Heterogeneous atypical cell populations are present in blood of metastatic breast cancer patients. Breast Cancer Res 16:R23

Showing the most recent 10 out of 87 publications