High gradient magnetic separation using ordered wire filters for the separation of human blood and bone marrow cells
High gradient magnetic separation using ordered wire filters for the separation of human blood and bone marrow cells
A novel conical funnel filter with an ordered wire array for the magnetic separation of fresh or previously cryopreserved human blood or bone marrow mononuclear cells has been constructed and tested. For the peripheral blood CD8+ mononuclear cell model 78 separations with 18 donors were done, 93% of the separations were done with previously cryopreserved cells. In the CD34++ bone marrow case, 15 separations with 7 donors were done, previously cryopreserved cells were used for 40% of the separations. Tetrameric antibody complexes were used to bind colloidal superparamagnetic dextran-iron particles to the cells with very low non-specific binding.
Ordered wire arrays with stainless steel wire from 50 to 150 μm diameter were constructed. To eliminate the distortion of the wire array with multiple use, supporting cross ties and crosslinking of the wires was used. The support and crosslinking of the wire array is believed to be novel. The hydrodynamic design of the overall filter was such that the retention of cells at zero field was on average 0.9%. The described hydrodynamic design and construction of the filter for separation application is believed to be novel.
For the CD8 model up to 109 total cells were used with no change in filter performance. After two successive separations an average purity of 98% was obtained. A third separation gave >99% purity.
It was observed that the use of electric AND magnetic fields on a single wire enabled sufficient induced charge on the cells of the Lorentz force to act strongly. This phenomenon in HGMS has not been previously described.
For the separation of CD34++ cells, suggestions for optimisation to ensure very high recoveries and purities were made.
University of Southampton
1993
Richards, Adrian John
(1993)
High gradient magnetic separation using ordered wire filters for the separation of human blood and bone marrow cells.
University of Southampton, Doctoral Thesis.
Record type:
Thesis
(Doctoral)
Abstract
A novel conical funnel filter with an ordered wire array for the magnetic separation of fresh or previously cryopreserved human blood or bone marrow mononuclear cells has been constructed and tested. For the peripheral blood CD8+ mononuclear cell model 78 separations with 18 donors were done, 93% of the separations were done with previously cryopreserved cells. In the CD34++ bone marrow case, 15 separations with 7 donors were done, previously cryopreserved cells were used for 40% of the separations. Tetrameric antibody complexes were used to bind colloidal superparamagnetic dextran-iron particles to the cells with very low non-specific binding.
Ordered wire arrays with stainless steel wire from 50 to 150 μm diameter were constructed. To eliminate the distortion of the wire array with multiple use, supporting cross ties and crosslinking of the wires was used. The support and crosslinking of the wire array is believed to be novel. The hydrodynamic design of the overall filter was such that the retention of cells at zero field was on average 0.9%. The described hydrodynamic design and construction of the filter for separation application is believed to be novel.
For the CD8 model up to 109 total cells were used with no change in filter performance. After two successive separations an average purity of 98% was obtained. A third separation gave >99% purity.
It was observed that the use of electric AND magnetic fields on a single wire enabled sufficient induced charge on the cells of the Lorentz force to act strongly. This phenomenon in HGMS has not been previously described.
For the separation of CD34++ cells, suggestions for optimisation to ensure very high recoveries and purities were made.
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Published date: 1993
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Local EPrints ID: 464486
URI: http://eprints.soton.ac.uk/id/eprint/464486
PURE UUID: 1fa84867-54f4-4564-89d5-bcd68c2e4641
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Date deposited: 04 Jul 2022 23:41
Last modified: 04 Jul 2022 23:41
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Author:
Adrian John Richards
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