Biomolecular gradients in cell culture systems

Keenan, Thomas; Folch, Albert

doi:10.1039/b711887b

Cited by 530 publications

(490 citation statements)

References 118 publications

(215 reference statements)

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“…It would be of considerable interest if these methods could be extended to the manipulation of solute concentration fields in microfluidic systems. Indeed, the ability to pattern and manipulate molecular concentration fields plays an essential role in several lab-on-a-chip applications and in controlled studies of biological processes such as development, inflammation, wound healing, and cancer, for which biomolecule gradients act as cellular signaling mechanisms [19]. The standard approach to precisely generate specified concentration gradients is to use microfluidic networks [20,21]-with limited temporal control, however.…”

Section: Introductionmentioning

confidence: 99%

Acoustic Tweezing and Patterning of Concentration Fields in Microfluidics

Karlsen

Bruus

2017

Phys. Rev. Applied

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We demonstrate theoretically that acoustic forces acting on inhomogeneous fluids can be used to pattern and manipulate solute concentration fields into spatiotemporally controllable configurations stabilized against gravity. A theoretical framework describing the dynamics of concentration fields that weakly perturb the fluid density and speed of sound is presented and applied to study manipulation of concentration fields in rectangular-channel acoustic eigenmodes and in Bessel-function acoustic vortices. In the first example, methods to obtain horizontal and vertical multilayer stratification of the concentration field at the end of a flow-through channel are presented. In the second example, we demonstrate acoustic tweezing and spatiotemporal manipulation of a local high-concentration region in a lower-concentration medium, thereby extending the realm of acoustic tweezing to include concentration fields.

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Section: Introductionmentioning

confidence: 99%

Acoustic Tweezing and Patterning of Concentration Fields in Microfluidics

Karlsen

Bruus

2017

Phys. Rev. Applied

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“…Its capacity is limited by poor gradient control and inability to visualize migrating cells (Keenan and Folch, 2008). Microfluidic devices, which can precisely configure gradient conditions, offer useful tools for cell migration and chemotaxis studies (Li and Lin, 2011;Wu et al, 2013).…”

Section: Introductionmentioning

confidence: 99%

Analysis of CCR7 mediated T cell transfectant migration using a microfluidic gradient generator

et al. 2015

Journal of Immunological Methods

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a b s t r a c t T lymphocyte migration is crucial for adaptive immunity. Manipulation of signaling molecules controlling cell migration combined with in vitro cell migration analysis provides a powerful research approach. Microfluidic devices, which can precisely configure chemoattractant gradients and allow quantitative single cell analysis, have been increasingly applied to cell migration and chemotaxis studies. However, there are a very limited number of published studies involving microfluidic migration analysis of genetically manipulated immune cells. In this study, we describe a simple microfluidic method for quantitative analysis of T cells expressing transfected chemokine receptors and other cell migration signaling probes. Using this method, we demonstrated chemotaxis of Jurkat transfectants expressing wild type or C terminus mutated CCR7 within a gradient of chemokine CCL19, and characterized the difference in transfectant migration mediated by wild type and mutant CCR7. The EGFP tagged CCR7 allows identification of CCR7 expressing transfectants in cell migration analysis and microscopy assessment of CCR7 dynamics. Collectively, our study demonstrated the effective use of the microfluidic method for studying CCR7 mediated T cell transfectant migration. We envision this developed method will provide a useful platform to functionally test various signaling mechanisms at the cell migration level.

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“…5 Microtechnologies offer unprecedented means to generate precise gradients of soluble and surface-tethered biomolecules, opening up exciting applications in (stem) cell biology. [6][7][8] For instance, microfluidic systems have been used to establish cytokine gradients to control human neural progenitor cell differentiation, 9 or to investigate the role of autocrine and paracrine signaling in regulating ESC self-renewal. 10 However, despite these exciting early examples, microfluidic gradient systems have not been widely used to address pertinent questions in stem cell biology.…”

Section: Introductionmentioning

confidence: 99%

Patterning of cell-instructive hydrogels by hydrodynamic flow focusing

Cosson

Allazetta

2013

Lab Chip

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Microfluidic gradient systems offer a very precise means to probe the response of cells to graded biomolecular signals in vitro, for example to model how morphogen proteins affect cell fate during developmental processes. However, existing gradient makers are designed for non-physiological plastic or glass cell culture substrates that are often limited in maintaining the phenotype and function of difficultto-culture mammalian cell types, such as stem cells. To address this bottleneck, we combine hydrogel engineering and microfluidics to generate tethered protein gradients on the surface of biomimetic poly(ethylene glycol) (PEG) hydrogels. Here we used software-assisted hydrodynamic flow focusing for exposing and rapidly capturing tagged proteins to gels in a step-wise fashion, resulting in immobilized gradients of virtually any desired shape and composition. To render our strategy amenable for highthroughput screening of multifactorial artificial cellular microenvironments, a dedicated microfluidic chip was devised for parallelization and multiplexing, yielding arrays of orthogonally overlapping gradients of up to 4 6 4 proteins. To illustrate the power of the platform for stem cell biology, we assessed how gradients of tethered leukemia inhibitory factor (LIF) influence embryonic stem cell (ESC) behavior. ESC responded to LIF gradients in a binary manner, maintaining the pluripotency marker Rex1/Zfp42 and forming self-renewing colonies above a threshold concentration of 85 ng cm 22. Our concept should be broadly applicable to probe how complex signaling microenvironments influence stem cell fate in culture.

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Biomolecular gradients in cell culture systems

Cited by 530 publications

References 118 publications

Acoustic Tweezing and Patterning of Concentration Fields in Microfluidics

Acoustic Tweezing and Patterning of Concentration Fields in Microfluidics

Analysis of CCR7 mediated T cell transfectant migration using a microfluidic gradient generator

Patterning of cell-instructive hydrogels by hydrodynamic flow focusing

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