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Three-dimensional tissue culture based on magnetic cell levitation

Abstract

Cell culture is an essential tool in drug discovery, tissue engineering and stem cell research. Conventional tissue culture produces two-dimensional cell growth with gene expression, signalling and morphology that can be different from those found in vivo, and this compromises its clinical relevance1,2,3,4,5. Here, we report a three-dimensional tissue culture based on magnetic levitation of cells in the presence of a hydrogel consisting of gold, magnetic iron oxide nanoparticles and filamentous bacteriophage. By spatially controlling the magnetic field, the geometry of the cell mass can be manipulated, and multicellular clustering of different cell types in co-culture can be achieved. Magnetically levitated human glioblastoma cells showed similar protein expression profiles to those observed in human tumour xenografts. Taken together, these results indicate that levitated three-dimensional culture with magnetized phage-based hydrogels more closely recapitulates in vivo protein expression and may be more feasible for long-term multicellular studies.

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Figure 1: Magnetic iron oxide-containing hydrogels.
Figure 2: Three-dimensional cell culture with magnetic-based levitation.
Figure 3: Comparison of three-dimensional cell growth with standard two-dimensional tissue culture.
Figure 4: Shape control of magnetically levitated culture.
Figure 5: Confrontation assay of magnetically levitated multicellular spheroids.

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Acknowledgements

The authors would like to thank R.R. Brentani, N.R. Pellis and E.H. Sage for helpful discussions and K. Dunner Jr and D. Bier for technical assistance. G.R.S. was supported by the Odyssey Scholar Program of the University of Texas M.D. Anderson Cancer Center and by the Breast Cancer Research Program of the US Department of the Defense (DOD). D.J.S. received support from the National Science Foundation. T.C.K. received support from the David and Lucille Packard Foundation. W.A. and R.P. received support from the Gillson-Longenbaugh Foundation, the Marcus Foundation, AngelWorks, DOD, National Institutes of Health (NIH) and National Cancer Institute.

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G.R.S., J.R.M., R.M.R., D.J.S., C.S.L, J.M., T.C.K., W.A. and R.P. conceived and designed the experiments. G.R.S., J.R.M., T.C.K., D.J.S., C.S.L., J.S.A. and J.M. performed the experiments. G.R.S., M.G.O., D.J.S., C.S.L., L.F.B., J.S.A., J.A.B., T.C.K., W.A. and R.P. analysed the data. R.M.R., M.M.G., J.A.B., J.G.G., T.C.K., W.A. and R.P. contributed materials and analysis tools. G.R.S., R.M.R., M.G.O., L.F.B., J.A.B., J.G.G., T.C.K., W.A. and R.P. co-wrote the paper.

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Correspondence to T. C. Killian, Wadih Arap or Renata Pasqualini.

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The University of Texas M. D. Anderson Cancer Center (UTMDACC) and Rice University (RU), along with their researchers, have filed patents on the technology and intellectual property reported here. If licensing or commercialization occurs, the researchers are entitled to standard royalties. G.R.S., R.M.R., C.S.L. and T.C.K. have equity in Nano3D Biosciences, Inc. UTMDACC and RU manage the terms of these arrangements in accordance to their established institutional conflict-of-interest policies.

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Souza, G., Molina, J., Raphael, R. et al. Three-dimensional tissue culture based on magnetic cell levitation. Nature Nanotech 5, 291–296 (2010). https://doi.org/10.1038/nnano.2010.23

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