Development of a shear stress-free microfluidic gradient generator capable of quantitatively analyzing single-cell morphology.
Microfluidics, the science of engineering fluid streams at the micrometer scale, offers unique tools for creating and controlling gradients of soluble compounds. Gradient generation can be used to recreate complex physiological microenvironments, but is also useful for screening purposes. For exampl...
| Publicado en: | Biomedical Microdevices Vol. 19; no. 4; pp. 1 - 11 |
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| Autores principales: | , , , , , , , |
| Formato: | Journal Article |
| Publicado: |
Springer Nature
Dec2017
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| Acceso en línea: | Ver este registro en EBSCOhost |
| fields | @attributes: recordID: 1 pdfLink: plink: https://search.ebscohost.com/login.aspx?direct=true&db=ccm&AN=125294504&site=ehost-live header: @attributes: shortDbName: ccm uiTerm: 125294504 longDbName: CINAHL Complete uiTag: AN controlInfo: bkinfo: dissinfo: jinfo: jid: 13872176 ODN jtl: Biomedical Microdevices issn: 13872176 maglogo: N pubinfo: dt: Dec2017 vid: 19 iid: 4 pid: 237 pub: Springer Nature place: New York, New York artinfo: ui: 125294504 10.1007/s10544-017-0222-z 125294504 ppf: 1 ppct: 10 formats: fmt: @attributes: type: P tig: atl: Development of a shear stress-free microfluidic gradient generator capable of quantitatively analyzing single-cell morphology. aug: au: Barata, David Spennati, Giulia Correia, Cristina Ribeiro, Nelson Harink, Björn Blitterswijk, Clemens Habibovic, Pamela Rijt, Sabine affil: Department of Tissue Regeneration, MIRA Institute for Biomedical Technology and Technical Medicine , University of Twente , 7500 AE Enschede The Netherlands sug: ab: Microfluidics, the science of engineering fluid streams at the micrometer scale, offers unique tools for creating and controlling gradients of soluble compounds. Gradient generation can be used to recreate complex physiological microenvironments, but is also useful for screening purposes. For example, in a single experiment, adherent cells can be exposed to a range of concentrations of the compound of interest, enabling high-content analysis of cell behaviour and enhancing throughput. In this study, we present the development of a microfluidic screening platform where, by means of diffusion, gradients of soluble compounds can be generated and sustained. This platform enables the culture of adherent cells under shear stress-free conditions, and their exposure to a soluble compound in a concentration gradient-wise manner. The platform consists of five serial cell culture chambers, all coupled to two lateral fluid supply channels that are used for gradient generation through a source-sink mechanism. Furthermore, an additional inlet and outlet are used for cell seeding inside the chambers. Finite element modeling was used for the optimization of the design of the platform and for validation of the dynamics of gradient generation. Then, as a proof-of-concept, human osteosarcoma MG-63 cells were cultured inside the platform and exposed to a gradient of Cytochalasin D, an actin polymerization inhibitor. This set-up allowed us to analyze cell morphological changes over time, including cell area and eccentricity measurements, as a function of Cytochalasin D concentration by using fluorescence image-based cytometry. pubtype: Academic Journal doctype: Journal Article ougenre: Article language: English refInfo: holdings: @attributes: islocal: N |
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