T cell chemotaxis in a simple microfluidic device

Lin, Francis; Butcher, Eugene C.

doi:10.1039/b607071j

Cited by 170 publications

(164 citation statements)

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“…The defined stable chemokine gradients are generated by controlled mixing of laminar flows of chemokines and medium. The mechanism for generating controlled stable chemical gradient in the "Y" device was characterized both experimentally and theoretically (Lin and Butcher, 2006). The cells were imaged~3 mm downstream of the "Y" junction.…”

Section: Pdms Microfluidic Device Preparation and Gradient Generationmentioning

confidence: 99%

“…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). Previously, we used a microfluidic gradient generating device to study the migration of activated human peripheral blood T cells (ahPBT) (Lin and Butcher, 2006;Nandagopal et al, 2011). Microfluidic devices were also used to study the migration of other subsets of human peripheral blood T cells such as memory T cells (Lin et al, 2008).…”

Section: Introductionmentioning

confidence: 99%

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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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Section: Pdms Microfluidic Device Preparation and Gradient Generationmentioning

confidence: 99%

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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“…Collectively, they highlight the complexity of the environmental regulation of NK-cell migrations in physiological and pathological conditions. NK-cell activation and functions are regulated by cytokine/ chemokine and/or DC in the microenvironments [18,[28][29][30] [36,37]. In this report, we demonstrated an application of such microfluidic device in examining how soluble factors produced by DC regulated NK-cell migrations in vitro.…”

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confidence: 99%

Microfluidic‐based, live‐cell analysis allows assessment of NK‐cell migration in response to crosstalk with dendritic cells

et al. 2014

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Migration and localization of NK cells into peripheral tissues are tightly regulated under normal and pathological conditions. The physiological importance of NK cell-DC crosstalk has been well documented. However, the ways in which DCs regulate the migratory properties of NK cells (such as chemotaxis, chemokinesis, chemo-repulsion) are not fully defined in vitro. Here, we employed a microfluidic platform to examine, at the single-cell level, C57BL/6 NK-cell migrations in a stable chemical gradient. We observed that soluble factors released by the immature and LPS-activated mature DCs induced a high level of chemotactic movement of IL-2-activated NK cells in vitro. We confirmed these findings in a standard trans-well migration assay, and identified CXCR3 as a key receptor on the NK cells that mediated the migration. More interestingly, we revealed a novel function of granulocyte macrophage colony-stimulating factor in repulsing NK-cell migrations. The future uses of such microfluidic device in the systematic evaluations of NK-cell migratory responses in NK cell-DC crosstalk will provide new insights into the development of DC-based NK-cell therapies against tumor and infections.Keywords: Bone marrow-derived dendritic cells r Chemo-repulsion r Chemotaxis r Microfluidic device r Natural killer cell r NK-DC crosstalk Additional supporting information may be found in the online version of this article at the publisher's web-site Introduction NK cells are motile bone marrow (BM)-derived lymphocytes that play a key role in innate immunity against viral, microbial infections, and transformed cells [1][2][3]. They are capable of killing transformed or infected cells [1,2,4,5], and/or producing cytokines/chemokines that can profoundly influence the quality and magnitude of the adaptive immune responses [6][7][8][9]. They Correspondence: Dr. Sam K. P. Kung e-mail: Sam.Kung2@med.umanitoba.ca acquire specific chemokine surface receptors during development and maturation [2,[10][11][12][13][14][15]. Chemokine receptors such as CCR7, CCR5, and CXCR3 are involved in the preferential migrations and localization of NK cells into the LNs [8,[16][17][18][19], whereas NK cells reside in blood, liver, and spleen exhibit higher CXCR1 and CX 3 CR1 expression [8,9,[20][21][22][23][24][25]. Nonchemokine family proteins such as chemerin and SIP 5 are also involved in the regulation of NK-cell trafficking [26,27]. Collectively, they highlight the complexity of the environmental regulation of NK-cell migrations in physiological and pathological conditions. NK-cell activation and functions are regulated by cytokine/ chemokine and/or DC in the microenvironments [18,[28][29][30] [36,37]. In this report, we demonstrated an application of such microfluidic device in examining how soluble factors produced by DC regulated NK-cell migrations in vitro. Results A microfluidic platform to perform live-cell imaging of NK-cell migrationsWe used the established Y-shaped microfluidic device to examine chemotactic or chemo-repulsive movements of NK cells ...

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“…Over each 0.8 mm length scale under our experimental conditions, the gradient prole was identical as characterized previously. 25 As shown in Fig. 2 (data from a different experiment than it in Fig.…”

Section: On-chip Selection Of Chemotactic Ascsmentioning

confidence: 86%

Selection of chemotactic adipose-derived stem cells using a microfluidic gradient generator

et al. 2015

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Vous avez des questions? Nous pouvons vous aider. Pour communiquer directement avec un auteur, consultez la première page de la revue dans laquelle son article a été publié afin de trouver ses coordonnées. Si vous n'arrivez pas à les repérer, communiquez avec nous à PublicationsArchive-ArchivesPublications@nrc-cnrc.gc.ca. Questions? Contact the NRC Publications Archive team atPublicationsArchive-ArchivesPublications@nrc-cnrc.gc.ca. If you wish to email the authors directly, please see the first page of the publication for their contact information. NRC Publications Archive Archives des publications du CNRCThis publication could be one of several versions: author's original, accepted manuscript or the publisher's version. / La version de cette publication peut être l'une des suivantes : la version prépublication de l'auteur, la version acceptée du manuscrit ou la version de l'éditeur. For the publisher's version, please access the DOI link below./ Pour consulter la version de l'éditeur, utilisez le lien DOI ci-dessous.http://doi.org/10.1039/C4RA12863JAccess and use of this website and the material on it are subject to the Terms and Conditions set forth at Selection of chemotactic adipose-derived stem cells using a microfluidic gradient generator Natarajan, Kanmani; Tian, Chantal; Xiang, Bo; Chi, Chao; Deng, Jixian; Zhang, Rundi; Freed, Darren H.; Arora, Rakesh C.; Tian, Ganghong; Lin, Francis http://nparc.cisti-icist.nrc-cnrc.gc.ca/fra/droits L'accès à ce site Web et l'utilisation de son contenu sont assujettis aux conditions présentées dans le site LISEZ CES CONDITIONS ATTENTIVEMENT AVANT D'UTILISER CE SITE WEB. NRC Publications Record / Notice d'Archives des publications de CNRC:http://nparc.cisti-icist.nrc-cnrc.gc.ca/eng/view/object/?id=3039e115-086c-46b1-89a1-b836cdb08b31 http://nparc.cisti-icist.nrc-cnrc.gc.ca/fra/voir/objet/?id=3039e115-086c-46b1-89a1-b836cdb08b31Selection of chemotactic adipose-derived stem cells using a microfluidic gradient generator † Kanmani Natarajan, ab Chantal Tian, a Bo Xiang, cd Chao Chi, def Jixian Deng, df Rundi Zhang, a Darren H. Freed, g Rakesh C. Arora, h Ganghong Tian * df and Francis Lin * abij Stem cells hold great promise for treating various degenerative diseases and conditions. However, the outcomes of preclinical and clinical cell therapy studies are still not close to our expectation. We believe that the unsatisfactory outcomes of cell therapy are at least partially due to insufficient homing of implanted stem cells into target organs and the use of heterogeneous cell populations for cell therapy.Therefore, there is a need to develop an effective guiding technique for stem cells to migrate to the target organs and to isolate effective stem cell populations. Toward this direction, we have previously demonstrated chemotaxis of rat adipose-derived stem cells (ASCs) to a well-defined gradient of epidermal growth factor (EGF) using a microfluidic device. In the current study, we further developed a microfluidics-based method for selecting chemotactic ASCs to EGF. This meth...

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T cell chemotaxis in a simple microfluidic device

Cited by 170 publications

References 32 publications

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

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

Microfluidic‐based, live‐cell analysis allows assessment of NK‐cell migration in response to crosstalk with dendritic cells

Selection of chemotactic adipose-derived stem cells using a microfluidic gradient generator

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