Enrichment of putative stem cells from adipose tissue using dielectrophoretic field-flow fractionation

Vykoukal, Jody; Vykoukal, Daynene; Freyberg, Susanne; Alt, Eckhard; Gascoyne, Peter R. C.

doi:10.1039/b717043b

Cited by 138 publications

(113 citation statements)

References 52 publications

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“…In more recent work, DEP has been used to obtain populations enriched for putative adult stem cells, as defined by expression of the stromal marker NG2 from enzyme-digested adipose tissue. 188 Of particular interest is the work of Flanagan et al, 189 whose objective was to determine whether stem cells and their more differentiated progeny can be identified by means other than flow cytometry. Mouse neural stem/precursor cells ͑NSPCs͒ and their differentiated derivatives ͑neurons and glia͒ were found to have distinct differences in their positive DEP behavior.…”

Section: Biomedicalmentioning

confidence: 99%

Review Article—Dielectrophoresis: Status of the theory, technology, and applications

2010

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A review is presented of the present status of the theory, the developed technology and the current applications of dielectrophoresis ͑DEP͒. Over the past 10 years around 2000 publications have addressed these three aspects, and current trends suggest that the theory and technology have matured sufficiently for most effort to now be directed towards applying DEP to unmet needs in such areas as biosensors, cell therapeutics, drug discovery, medical diagnostics, microfluidics, nanoassembly, and particle filtration. The dipole approximation to describe the DEP force acting on a particle subjected to a nonuniform electric field has evolved to include multipole contributions, the perturbing effects arising from interactions with other cells and boundary surfaces, and the influence of electrical double-layer polarizations that must be considered for nanoparticles. Theoretical modelling of the electric field gradients generated by different electrode designs has also reached an advanced state. Advances in the technology include the development of sophisticated electrode designs, along with the introduction of new materials ͑e.g., silicone polymers, dry film resist͒ and methods for fabricating the electrodes and microfluidics of DEP devices ͑photo and electron beam lithography, laser ablation, thin film techniques, CMOS technology͒. Around three-quarters of the 300 or so scientific publications now being published each year on DEP are directed towards practical applications, and this is matched with an increasing number of patent applications. A summary of the US patents granted since January 2005 is given, along with an outline of the small number of perceived industrial applications ͑e.g., mineral separation, micropolishing, manipulation and dispensing of fluid droplets, manipulation and assembly of micro components͒. The technology has also advanced sufficiently for DEP to be used as a tool to manipulate nanoparticles ͑e.g., carbon nanotubes, nano wires, gold and metal oxide nanoparticles͒ for the fabrication of devices and sensors. Most efforts are now being directed towards biomedical applications, such as the spatial manipulation and selective separation/enrichment of target cells or bacteria, high-throughput molecular screening, biosensors, immunoassays, and the artificial engineering of three-dimensional cell constructs. DEP is able to manipulate and sort cells without the need for biochemical labels or other bioengineered tags, and without contact to any surfaces. This opens up potentially important applications of DEP as a tool to address an unmet need in stem cell research and therapy.

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Section: Biomedicalmentioning

confidence: 99%

Review Article—Dielectrophoresis: Status of the theory, technology, and applications

2010

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“…7 Several types of stem and progenitor cells (hematopoietic, adipose-derived progenitors, and myoblasts) have been successfully analyzed by flow cytometry after DEP sorting. [7][8][9]11 Furthermore, DEP sorted myoblasts have high viability post-sorting. 11 Mouse NSPCs isolated by DEP were analyzed in differentiation assays, demonstrating that the cells retained the ability to generate neurons and astrocytes post-DEP sorting.…”

Section: Ease Of Fabrication and Usementioning

confidence: 99%

“…To further simplify fabrication of the electrode array, a commercially available flexible circuit could be purchased. 9 Operation of the device is also simple as all that is needed is a compressed gas tank (already available in many biology and medical research labs) with a pressure regulator and a waveform generator. A hemostat performs the function of a valve and enables reliable switching of fluids if pneumatic or digitally controlled fluid valves are not available.…”

Section: Ease Of Fabrication and Usementioning

confidence: 99%

“…10,29 Other goals have been to design DEP devices for clinical samples (needing eventual approval by the FDA) or to make them amenable to industrial scale up or mass production. 9,24,27 However, many labs are recognizing the need to simplify the operation of DEP sorters to make them more useful for biological separations. To this end, new generations of sorters have addressed issues such as improving cell recovery from DEP devices.…”

Section: Ease Of Fabrication and Usementioning

confidence: 99%

“…5 These include stem cells from blood or tissue-CD34-positive hematopoietic stem cells have been enriched from bone marrow or peripheral blood 7,8 and NG2-positive human adipose progenitor cells were enriched 14-fold from tissue. 9 DEP-based separation can isolate undifferentiated from more differentiated cells in the same lineage, as shown by the separation of neural stem and progenitor cells (NSPCs) from differentiated neurons 10 and separation of C2C12 myoblasts and more differentiated myotubes. 11 Progenitor cells within the same lineage are also amenable to separation using DEP, and enrichment of neuron progenitors and astrocyte progenitors from a mixed population of NSPCs by DEP provides significantly better enrichment than FACS sorting with PSA-NCAM-a purported marker for neuron progenitors.…”

Section: Introduction/backgroundmentioning

confidence: 99%

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Increasing label-free stem cell sorting capacity to reach transplantation-scale throughput

et al. 2014

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Dielectrophoresis (DEP) has proven an invaluable tool for the enrichment of populations of stem and progenitor cells owing to its ability to sort cells in a labelfree manner and its biological safety. However, DEP separation devices have suffered from a low throughput preventing researchers from undertaking studies requiring large numbers of cells, such as needed for cell transplantation. We developed a microfluidic device designed for the enrichment of stem and progenitor cell populations that sorts cells at a rate of 150,000 cells/h, corresponding to an improvement in the throughput achieved with our previous device designs by over an order of magnitude. This advancement, coupled with data showing the DEPsorted cells retain their enrichment and differentiation capacity when expanded in culture for periods of up to 2 weeks, provides sufficient throughput and cell numbers to enable a wider variety of experiments with enriched stem and progenitor cell populations. Furthermore, the sorting devices presented here provide ease of setup and operation, a simple fabrication process, and a low associated cost to use that makes them more amenable for use in common biological research laboratories. To our knowledge, this work represents the first to enrich stem cells and expand them in culture to generate transplantation-scale numbers of differentiation-competent cells using DEP. V C 2014 AIP Publishing LLC.

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Dielectrophoretic Studies of Bioparticles

2017

Dielectrophoresis

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Enrichment of putative stem cells from adipose tissue using dielectrophoretic field-flow fractionation

Cited by 138 publications

References 52 publications

Review Article—Dielectrophoresis: Status of the theory, technology, and applications

Review Article—Dielectrophoresis: Status of the theory, technology, and applications

Increasing label-free stem cell sorting capacity to reach transplantation-scale throughput

Dielectrophoretic Studies of Bioparticles

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