Magnetic Flow Sorting Using a Model System of Human Lymphocytes and a Colloidal Magnetic Label

Zborowski, Maciej; Moore, Lee R.; Reddy, Susheel; Chen, Guo Hua; Sun, Liping; Chalmers, Jeffrey J.

doi:10.1097/00002480-199609000-00071

Cited by 13 publications

(9 citation statements)

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“…Earlier DMF prototypes were tested for fractionation of immunomagnetically labeled, peripheral blood T-lymphocytes and an immortalized T cell line (Jurkat). 30,31,53 Compared to T-lymphocytes, the fractionation of the KG-1a cell line presents a more challenging model because of the lower expression of the CD34 cell surface antigen on KG-1a cells as compared to the CD45 antigen expression of the T-lymphocytes. [54][55][56] This results in lower KG-1a cell magnetophoretic mobilities.…”

Section: Discussionmentioning

confidence: 99%

Sequential CD34 cellfractionation by magnetophoresis in a magnetic dipole flow sorter

Schneider

Karl

Moore

et al. 2010

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Cell separation and fractionation based on fluorescent and magnetic labeling procedures are common tools in contemporary research. These techniques rely on binding of fluorophores or magnetic particles conjugated to antibodies to target cells. Cell surface marker expression levels within cell populations vary with progression through the cell cycle. In an earlier work we showed the reproducible magnetic fractionation (single pass) of the Jurkat cell line based on the population distribution of CD45 surface marker expression. Here we present a study on magnetic fractionation of a stem and progenitor cell (SPC) population using the established acute myelogenous leukemia cell line KG-1a as a cell model. The cells express a CD34 cell surface marker associated with the hematopoietic progenitor cell activity and the progenitor cell lineage commitment (related to the CD34 marker expression level). The CD34 expression level is approximately an order of magnitude lower than that of the CD45 marker, which required further improvements of the magnetic fractionation apparatus. The cells were immuno-magnetically labeled using a sandwich of anti CD34 antibody-phycoerythrin (PE) conjugate and anti PE magnetic nanobead and fractionated into eight components using a continuous flow dipole magnetophoresis apparatus. The CD34 marker expression distribution between sorted fractions was measured by quantitative PE flow cytometry (using QuantiBRITE™ PE calibration beads), and it was shown to be correlated with the cell magnetophoretic mobility distribution. A flow outlet addressing scheme based on the concept of the transport lamina thickness was used to control cell distribution between the eight outlet ports. The fractional cell distributions showed good agreement with numerical simulations of the fractionation based on the cell magnetophoretic mobility distribution in the unsorted sample.

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Section: Discussionmentioning

confidence: 99%

Sequential CD34 cellfractionation by magnetophoresis in a magnetic dipole flow sorter

Schneider

Karl

Moore

et al. 2010

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“…A previously published, two-step immunofluoromagnetic labeling procedure was employed (21). This procedure allows for both magnetic separation and flow cytometry analysis.…”

Section: Immunomagnetic Labeling Of Lymphocytesmentioning

confidence: 99%

Continuous, flow-through immunomagnetic cell sorting in a quadrupole field

et al. 1998

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A flow-through quadrupole magnetic cell separator has been designed, built, and evaluated by using a cell model system of human peripheral T lymphocytes (CD4 ϩ , CD8 ϩ , and CD45 ϩ cells). The immunomagnetic labeling was accomplished by using a sandwich of mouse anti-human monoclonal antibody conjugated to fluorescein isothiocyanate and rat anti-mouse polyclonal antibody conjugated to a colloidal magnetic nanoparticle. The feed and sorted fractions were analyzed by FACScan flow cytometry. The magnetically labeled cells were separated from nonlabeled ones in a flow-through cylindrical column within a quadrupole field, which exerted a radial, outward force on the magnetic cells. The flow rate of the cell samples was 0.1-0.75 ml/min, and the flow rate of sheath fluid was 1.5-33.3 times that of the sample flow rate. The maximum shear stress exerted on the cell was less than 1 dyne/cm 2 , which was well below the level that would threaten cell integrity and membrane disruption. The maximum magnetic field was 0.765 T at the channel wall, and the gradient was 0.174 T/mm. The highest purity of selected cells was 99.6% (CD8 cells, initial purity of 26%), and the highest recovery of selected cells was 79% (CD4 cells, initial purity of 20%). The maximum throughput of the quadrupole magnetic cell separator was 7,040 cells/s (CD45 cells, initial purity of 5%). Theoretical calculations showed that the throughput can be increased to 10 6 cells/s by a scale-up of the current prototype. Cytometry 33: 469-475, 1998.

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“…Magnetic micro/nanoparticle-based cell sorting is a second common approach that utilizes antibody specificity to separate cells of interest from bulk cell mixtures and is often inexpensive and easy to use. ,, However, magnetic interaction between microparticles and some sensitive cell populations, such as stem and progenitor cells, is known to have a negative impact on cell viability, phenotypic identity, and cell function. − Furthermore, the high concentrations of small particles (<2 μm) commonly used in magnetic isolation risk significant cell stress due to particle internalization . Magnetic cell sorting has been shown to introduce false positives largely due to nonspecific cell–particle adhesion . Adhesion-based cell isolation approaches, especially those combined with several microfluidic separation technologies (differential adhesion, hierarchical nanosubstrates, negative selection, size-based separation, and dielectrophoresis, etc.)…”

Section: Introductionmentioning

confidence: 99%

Cell Isolation and Recovery Using Hollow Glass Microspheres Coated with Nanolayered Films for Applications in Resource-Limited Settings

Dong

Ahrens

et al. 2017

ACS Appl. Mater. Interfaces

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Established cell isolation and purification techniques such as fluorescence-activated cell sorting (FACS), isolation through magnetic micro/nanoparticles, and recovery via microfluidic devices have limited application as disposable technologies appropriate for point-of-care use in remote areas where lab equipment as well as electrical, magnetic, and optical sources are restricted. We report a simple yet effective method for cell isolation and recovery that requires neither specialized lab equipment nor any form of power source. Specifically, self-floating hollow glass microspheres were coated with an enzymatically degradable nanolayered film and conjugated with antibodies to allow both fast capture and release of subpopulations of cells from a cell mixture. Targeted cells were captured by the microspheres and allowed to float to the top of the hosting liquid, thereby isolating targeted cells. To minimize nonspecific adhesion of untargeted cells and to enhance the purity of the isolated cell population, an antifouling polymer brush layer was grafted onto the nanolayered film. Using the EpCAM-expressing cancer cell line PC-3 in blood as a model system, we have demonstrated the isolation and recovery of cancer cells without compromising cell viability or proliferative potential. The whole process takes less than 1 h. To support the rational extension of this platform technology, we introduce extensive characterization of the critical design parameters: film formation and degradation, grafting with a poly(ethylene glycol) (PEG) sheath, and introducing functional antibodies. Our approach is expected to overcome practical hurdles and provide viable targeted cells for downstream analyses in resource-limited settings.

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Magnetic Flow Sorting Using a Model System of Human Lymphocytes and a Colloidal Magnetic Label

Cited by 13 publications

References 0 publications

Sequential CD34 cellfractionation by magnetophoresis in a magnetic dipole flow sorter

Sequential CD34 cellfractionation by magnetophoresis in a magnetic dipole flow sorter

Continuous, flow-through immunomagnetic cell sorting in a quadrupole field

Cell Isolation and Recovery Using Hollow Glass Microspheres Coated with Nanolayered Films for Applications in Resource-Limited Settings

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