ArticleExperimental hematology2007
A novel high throughput immunomagnetic cell sorting system for potential clinical scale depletion of T cells for allogeneic stem cell transplantation.
Article in Experimental hematology, 2007. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed, 21 citations in OpenAlex.
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- Detection of Rare Objects by Flow Cytometry: Imaging, Cell Sorting, and Deep Learning Approaches.International journal of molecular sciences · 2020Review
- Optofluidic device for the quantification of circulating tumor cells in breast cancer.Scientific reports · 2017Article
- Fundamentals and application of magnetic particles in cell isolation and enrichment: a review.Reports on progress in physics. Physical Society (Great Britain) · 2015Review
- Circulating tumor cells: a review of present methods and the need to identify heterogeneous phenotypes.Annals of clinical and laboratory science · 2013Review
- Simultaneous, single particle, magnetization and size measurements of micron sized, magnetic particles.Journal of magnetism and magnetic materials · 2012Article
- Emerging technologies for CTC detection based on depletion of normal cells.Recent results in cancer research. Fortschritte der Krebsforschung. Progres dans les recherches sur le cancer · 2012Review
- Computational Fluid Dynamics Simulation of a Quadrupole Magnetic Sorter Flow Channel: Effect of Splitter Position on Nonspecific Crossover.The Canadian journal of chemical engineering · 2011Article
- Manipulation of magnetically labeled and unlabeled cells with mobile magnetic traps.Biophysical journal · 2010Article
- Magnetic wire traps and programmable manipulation of biological cells.Physical review letters · 2009Article
- Quadrupole magnetic sorting of porcine islets of Langerhans.Tissue engineering. Part C, Methods · 2009Article
- Optimization of an enrichment process for circulating tumor cells from the blood of head and neck cancer patients through depletion of normal cells.Biotechnology and bioengineering · 2009Article
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Authors and funding
5 authors at 3 institutions in 1 country.
Funding
Abstract
objectiveTo develop an immunomagnetic cell separation system for allogeneic hematopoietic stem cell (HSC) transplantations, which can achieve a high level of T-cell depletion (at least 4.0 log(10)), high level of recovery of hematopoietic stem cells (>90%), with a high throughput (>10(6) cells/second).
methodsPeripheral blood leukocytes (PBLs) from buffy coats were spiked with CD34-expressing cells (KG1a) to mimic a leukaphoresis product containing stimulated HSCs. T cells were labeled with anti-CD3(+) Dynabeads and separated in a quadrupole magnetic cell sorter (QMS). The performance of the system with respect to T-cell depletion and recovery of non-T cells and spiked KG1a was determined using four-color, flow cytometry analysis, with the aid of Trucount cell-concentration calibration beads. Limiting dilution assays were also performed to quantify the log(10) depletion of clonable T cells.
resultsWhile the general performance of the QMS system is governed by proven theoretical principles, significant system variability exist, not all of which can be explained by our current understanding. Consequently, a factorial design was employed, guided by JMP software, to optimize the labeling conditions and operation of the QMS focused on maximizing the depletion of T cell, and recovery of unlabeled cells including KG1a cells. From these studies, an optimized, no wash, immunomagnetic labeling protocol and optimized QMS operating conditions were developed. For an average initial cell concentration of 1.7 x 10(8) total cells, an average 3.96 +/- 0.33 log(10) depletion (range, 3.53-4.34) of CD3(+)CD45(+) cells with a mean 99.43% +/- 4.23% recovery of CD34(+)CD45(+) cells (range, 94.38-104.90%) was achieved at a sorting speed of 10(6) cells/s (n = 6). Limiting dilution assays on the T-cell depleted fractions, which gave a log(10) depletion of 3.51 for the clonable T cells.
conclusionWe suggest that this system will provide superior performance with respect to T-cell depletion and CD34(+) recovery for clinical allogeneic bone marrow transplants. Ongoing studies, on a clinical scale, are being conducted to demonstrate this claim.
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