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Merck

Rapid cell separation with minimal manipulation for autologous cell therapies.

Scientific reports (2017-02-06)
Alban J Smith, Richard D O'Rorke, Akshay Kale, Roberts Rimsa, Matthew J Tomlinson, Jennifer Kirkham, A Giles Davies, Christoph Wälti, Christopher D Wood
RÉSUMÉ

The ability to isolate specific, viable cell populations from mixed ensembles with minimal manipulation and within intra-operative time would provide significant advantages for autologous, cell-based therapies in regenerative medicine. Current cell-enrichment technologies are either slow, lack specificity and/or require labelling. Thus a rapid, label-free separation technology that does not affect cell functionality, viability or phenotype is highly desirable. Here, we demonstrate separation of viable from non-viable human stromal cells using remote dielectrophoresis, in which an electric field is coupled into a microfluidic channel using shear-horizontal surface acoustic waves, producing an array of virtual electrodes within the channel. This allows high-throughput dielectrophoretic cell separation in high conductivity, physiological-like fluids, overcoming the limitations of conventional dielectrophoresis. We demonstrate viable/non-viable separation efficacy of >98% in pre-purified mesenchymal stromal cells, extracted from human dental pulp, with no adverse effects on cell viability, or on their subsequent osteogenic capabilities.

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Leukocyte Alkaline Phosphatase Kit, based on naphthol AS-MX phosphate and fast blue RR salt