Cell separation mediated by differential rolling adhesion

Greenberg, Adam W.; Hammer, Daniel A.

doi:10.1002/bit.1043

Cited by 39 publications

(48 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Furthermore, cells near the edge rolled at an average velocity of 1 µm/s, whereas cells away from the edge rolled at an average velocity of 0.5 µm/s. These results demonstrate that the P-selectin edge enabled control over the transport of rolling cells by (a) changing the direction of rolling, and (b) increasing the rolling speed.Similar experiments were performed with Sialyl Lewis(x) (sLe x ) coated microspheres that form transient bonds with P-selectin and are used as models to study cell rolling 22,23,31 . The microspheres rolled selectively on the P-selectin region with average velocity of about 3-4 µm/s at a flow speed of 200 µL/min corresponding to a shear stress of 0.33 dyn/cm 2 (0.033 Pa).…”

mentioning

confidence: 99%

“…Cell rolling has been extensively studied due to its important role in physiological processes such as recruitment of leukocytes to sites of inflammation, homing of hematopoietic progenitor cells after intravenous injection, tumor cell metastasis and other inflammatory processes [19][20][21] . Cell rolling has been achieved in vitro and mimicked using microsphere models 22,23 . Researchers have also investigated the possibility of separating cells based on rolling, including a technique for capturing cells in microfluidic devices 24,25 and separation of cells based on differential rolling velocities in a flow chamber 23 .…”

mentioning

confidence: 99%

“…This unique technique may facilitate the incorporation of factors such as cell deformability, morphology and receptor clustering that influence cell rolling, but are typically inaccessible to conventional techniques of cell separation 29,30,34,35 , while simultaneously enabling the degree of specificity to be tuned through co-immobilization of antibodies to specific cell markers 26 . With recent research uncovering the links between nanomechanical characteristics of cells and properties such as states of disease 36,37 and homing ability of cancer cells and stem cells 23 , the development of separation techniques based on nanomechanical control of cell rolling through patterning of receptors that depends on both the mechanical properties of cells as well as specific receptor-ligand interactions may prove to be a very useful method for separation and analysis of stem cells, cancer cells, and white blood cells.…”

mentioning

confidence: 99%

See 2 more Smart Citations

Nanomechanical Control of Cell Rolling in Two Dimensions through Surface Patterning of Receptors

et al. 2008

View full text Add to dashboard Cite

We envisioned that label-free control of the transport of cells in two dimensions through receptorligand interactions would enable simple separation systems that are easy to implement, yet retain the specificity of receptor-ligand interactions. Here we demonstrate nanomechanical control of cell transport in two dimensions via transient receptor-ligand adhesive bonds by patterning of receptors that direct cell rolling through an edge effect. HL-60 cells rolling on P-selectin receptor patterns were deflected at angles of 5 -10° with respect to their direction of travel. Absence of this effect in the case of rigid microsphere models of cell rolling suggests that this twodimensional motion depends on nanomechanical properties of the rolling cell. This work suggests the feasibility of simple continuous-flow microfluidic cell separation systems that minimize processing steps and yet retain the specificity of receptor-ligand interactions.Techniques for separation of cells rely on the ability to control their transport based on specific characteristics such as size, density, or surface ligands. In particular, control of transport in two dimensions is highly advantageous for the design of continuous-flow separation systems [1][2][3] . Continuous-flow separation of cells based on specific receptor-ligand interactions is currently limited to external control techniques that harness dielectrophoretic, magnetic, or other forces 4, 5 and typically rely on capturing or labeling of the cells. These label-based approaches are very useful for cell separation, but involve special apparatus and multiple processing steps for labeling and label removal which are not desirable for sensitive cell samples or for point of care diagnostics, especially in third world countries. On the other hand, label-free approaches based on physical characteristics of the cells are often easy to implement, but lack the specificity provided by receptor-ligand interactions 4 . Control over the transport of cells based on specific receptor-ligand interactions without labeling and label-removal steps would enable cell separation devices for single use or continuous-flow separation, while retaining the specificity of receptor-ligand interactions. In this paper, we wanted to examine whether receptor patterning could be used to achieve nanomechanical control of the transport of cells in two dimensions in a label-free manner through the formation of transient receptor-ligand bonds that result in cell rolling.The formation of transient receptor-ligand bonds commonly occurs between cells flowing in the blood stream and the vascular endothelium in a physiological process known as cell rolling [6][7][8] . This phenomenon is mediated primarily by glycoprotein receptors known as selectins, among some other receptors can also enable cell rolling [8][9][10][11] . The adhesive bonds formed by selectins have high dissociation rates and are also responsive to shear stress [12][13][14] .As the cell rolls under shear force exerted by the flowing blood stream, bonds ...

show abstract

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

Nanomechanical Control of Cell Rolling in Two Dimensions through Surface Patterning of Receptors

et al. 2008

View full text Add to dashboard Cite

show abstract

“…This ability may enable higher resolution for distinguishing between cells with different rolling characteristics at the single cell level. As cell rolling behavior depends on the ligands on the cell, we hypothesize that such experiments may enable design of appropriate patterns to separate different cell phenotypes based on differences in their rolling behavior 3,[9][10][11][12] . This work can thus be useful for design of devices for cell separation and analysis based on interaction between cell ligands and the patterned asymmetric receptors.…”

Section: Discussionmentioning

confidence: 99%

Studying Cell Rolling Trajectories on Asymmetric Receptor Patterns

Lee

Bose

Vliet

et al. 2011

JoVE

View full text Add to dashboard Cite

Lateral displacement of cells orthogonal to a flow stream by rolling on asymmetric receptor patterns presents an opportunity for development of new devices for label-free separation and analysis of cells 1 . Such devices may use lateral displacement for continuous-flow separation, or receptor patterns that modulate adhesion to distinguish between different cell phenotypes or levels of receptor expression. Understanding the nature of cell rolling trajectories on receptor-patterned substrates is necessary for engineering of the substrates and design of such devices.Here, we demonstrate a protocol for studying cell rolling trajectories on asymmetric receptor patterns that support cell rolling adhesion 2 . Well-defined, μm-scale patterns of P-selectin receptors were fabricated using microcontact printing on gold-coated slides that were incorporated in a flow chamber. HL60 cells expressing the PSGL-1 ligand 3 were flowed across a field of patterned lines and visualized on an inverted bright field microscope. The cells rolled and tracked along the inclined edges of the patterns, resulting in lateral deflection 1 . Each cell typically rolled for a certain distance along the pattern edges (defined as the edge tracking length), detached from the edge, and reattached to a downstream pattern. Although this detachment makes it difficult to track the entire trajectory of a cell from entrance to exit in the flow chamber, particle-tracking software was used to analyze and yield the rolling trajectories of the cells during the time when they were moving on a single receptor-patterned line. The trajectories were then examined to obtain distributions of cell rolling velocities and the edge tracking lengths for each cell for different patterns.This protocol is useful for quantifying cell rolling trajectories on receptor patterns and relating these to engineering parameters such as pattern angle and shear stress. Such data will be useful for design of microfluidic devices for label-free cell separation and analysis. Using microcontact printing (μCP) 4-7 to make alternating self-assembled monolayers (SAMs) of PEG molecules on the gold-coated glass slides: Fabricate microcontact printing polydimethylsiloxane (PDMS) stamps that defined the receptor patterns with inclination angle of α = 10°by an SU-8 molding process. Clean the gold surface with piranha solution (3:1 mixture of sulfuric acid to 30% hydrogen peroxide) for 20 minutes and then rinse the surface with copious DI water at 24.5°C prior to use. Ink the PDMS stamp with 5mM PEG solution in ethanol. Dry the stamp in air for 20 minutes. Gently put the stamp on the gold surface for 40 sec and make sure there is a good contact between the gold surface and the stamp. No excess pressure is required. Rinse the surface with ethanol and dry it under a stream of N2. 2. Incubate the substrate within P-selectin solution (15 μg/mL in DPBS) using a perfusion chamber (Electron Microscopy Sciences) for 3 hours at 24.5°C to pattern the remaining areas with P-selectin. Rinse the surface w...

show abstract

“…29 To silence genes in circulating cells, an implantable device for siRNA delivery was developed in this study. On the basis of our previous results, 18,19,25,30 P-selectin, an adhesion molecule generated in endothelial and platelet cells and involved in the capture of leukocytes from the bloodstream, 22,31 was selected to construct liposome-based targeted NPs for siRNA delivery.…”

Section: Discussionmentioning

confidence: 99%

An immobilized nanoparticle-based platform for efficient gene knockdown of targeted cells in the circulation

Huang

King

2009

Gene Ther

View full text Add to dashboard Cite

It is well established that specific interaction between adhesion molecules of endothelial cells and receptors on leukocytes can separate and recruit leukocytes from the bloodstream to sites of inflammation and coagulation. Previously, we showed that P-selectin can be absorbed onto the surface of a bloodcompatible microrenathane tube, and the P-selectin-coated surface could successfully capture P-selectin receptor-positive stem cells from physiological shear flow in vitro and from the bloodstream in vivo. In this paper, P-selectin was covalently attached to the surface of nanoscale liposomes to create targeting nanoparticles (NPs). Small interfering RNA (siRNA) was encapsulated by these nanoscale liposomes, and the liposomes were stabilized by PEGylation with DSPE-PEG2000. Experiments showed that these P-selectin-, PEGylated-, nanoscale-liposomes (PS-DSPE-PEG NPs) could be absorbed onto the inner surface of microrenathane tubing. The coated surface could specifically capture targeted cells from physiological shear flow, efficiently deliver encapsulated siRNA into adherent cells and dramatically silence the targeted gene neutrophil elastase. With this device, we create a high localized concentration for siRNA delivery in the circulatory system, providing circulating target cells adequate time to interact with therapeutic materials. SiRNA is efficaciously delivered into specific target cells, thereby providing a powerful tool for highly efficient siRNA transfection and other therapeutic materials delivery in circulation. The method should prove especially useful for diseases derived from disorders of blood cells.

show abstract

Cell separation mediated by differential rolling adhesion

Cited by 39 publications

References 30 publications

Nanomechanical Control of Cell Rolling in Two Dimensions through Surface Patterning of Receptors

Nanomechanical Control of Cell Rolling in Two Dimensions through Surface Patterning of Receptors

Studying Cell Rolling Trajectories on Asymmetric Receptor Patterns

An immobilized nanoparticle-based platform for efficient gene knockdown of targeted cells in the circulation

Contact Info

Product

Resources

About