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Material surface engineering for multiplex cell culture in microwell

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Abstract

In this study, we develop a new concept for multiplexed and localized cell co-culture. This cell chip consists of a polystyrene spin-coated solid support bearing gold-bottomed microwells. The cell-chip support is fabricated as follows: (i) electrosputtering of a thin layer of gold (40 nm) onto a polycarbonate substrate, (ii) spin coating of a polystyrene thin film (500 ± 50 nm) over the gold layer, followed by (iii) polystyrene etching through the spotting of toluene nanovolume (300–900 pL). In each gold-bottomed microwell, a small population of adherent cells (approx. 100 cells) can be cultured. In this miniaturized system, different cell lines can be co-cultured on a 1-cm2 surface, opening the way to multiplexed cell-chip development. In order to keep the cells in a properly hydrated environment and to physically retain them before they adhere, a biocompatible alginate polymer was used during the robotized micropipetting. This approach allows for the encapsulation of the cell in a very small volume (50 nL), directly in the microwells. After 24 h of culture, the cells adhered on the gold bottom of the microwells, and the alginate matrix was removed by addition of calcium-free culture medium.

Graphical Abstract

Multiplex culture of cells was obtained using in situ produced microwells and encapsulated cells. The microwells are produced by organic solvent etching (nanovolume spotting) of a spin-coated polystyrene thin film, and the living multiple cell line deposition is obtained using on-site encapsulation in an alginate bead.

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Authors and Affiliations

  1. Laboratoire de Génie Enzymatique, Membranes Biomimétiques et Assemblages Supramoléculaires, Institut de Chimie et Biochimie Moléculaires et Supramoléculaires, Université Claude Bernard Lyon 1 – University of Lyon – CNRS 5246 ICBMS, Bât. CPE – 43 Bd du 11 Nov. 1918, 69622, Villeurbanne, France

    Ophélie I. Berthuy, Guillaume G. Octobre, Loïc J. Blum & Christophe A. Marquette

  2. AXO Science, 66 Boulevard Niels Bohr, 69100, Villeurbanne, France

    Céline A. Mandon & Benjamin P. Corgier

  3. Européen Commission, Joint Research Centre, Institute for Heath and Consumer Protection, Ispra, VA, Italy

    Giacomo Ceccone & Valentina Spampinato

Authors
  1. Ophélie I. Berthuy
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  2. Céline A. Mandon
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  3. Benjamin P. Corgier
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  4. Guillaume G. Octobre
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  5. Giacomo Ceccone
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  6. Valentina Spampinato
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  7. Loïc J. Blum
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  8. Christophe A. Marquette
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Correspondence to Christophe A. Marquette.

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Berthuy, O.I., Mandon, C.A., Corgier, B.P. et al. Material surface engineering for multiplex cell culture in microwell. J Mater Sci 49, 4481–4489 (2014). https://doi.org/10.1007/s10853-014-8145-z

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  • DOI: https://doi.org/10.1007/s10853-014-8145-z

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