Material surface engineering for multiplex cell culture in microwell
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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.
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.
KeywordsAlginate Bare Gold Alginate Capsule Microwell Array Microspectroscopy Analysis
- 14.Lecault V, Vaninsberghe M, Sekulovic S, Knapp DJ, Wohrer S, Bowden W, Viel F, McLaughlin T, Jarandehei A, Miller M, Falconnet D, White AK, Kent DG, Copley MR, Taghipour F, Eaves CJ, Humphries RK, Piret JM, Hansen CL (2011) High-throughput analysis of single hematopoietic stem cell proliferation in microfluidic cell culture arrays. Nat Methods 8(7):581–586CrossRefGoogle Scholar
- 18.Beamson G, Briggs D (1992) The XPS of polymer database. Surface Spectra, ChichesterGoogle Scholar
- 19.Vickerman JC, Briggs D (2001) ToF–SIMS: materials analysis by mass spectrometry, 2nd edn. Surface Spectra, ChichesterGoogle Scholar