Metre-long cell-laden microfibres exhibit tissue morphologies and functions

Abstract

Artificial reconstruction of fibre-shaped cellular constructs could greatly contribute to tissue assembly in vitro. Here we show that, by using a microfluidic device with double-coaxial laminar flow, metre-long core–shell hydrogel microfibres encapsulating ECM proteins and differentiated cells or somatic stem cells can be fabricated, and that the microfibres reconstitute intrinsic morphologies and functions of living tissues. We also show that these functional fibres can be assembled, by weaving and reeling, into macroscopic cellular structures with various spatial patterns. Moreover, fibres encapsulating primary pancreatic islet cells and transplanted through a microcatheter into the subrenal capsular space of diabetic mice normalized blood glucose concentrations for about two weeks. These microfibres may find use as templates for the reconstruction of fibre-shaped functional tissues that mimic muscle fibres, blood vessels or nerve networks in vivo.

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Figure 1: Formation of a metre-long cell-laden microfibre.
Figure 2: Intrinsic cellular morphologies and functions of cell fibres using primary cells.
Figure 3: Differentiation induction of primary NSC–PCol fibres.
Figure 4: Fibre-based assembly of higher-order 3D macroscopic cellular structures.
Figure 5: Transplantation of primary islet cell fibres into diabetic mice.

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Acknowledgements

We thank S. Seino (Kobe University) for providing the MIN6m9 cell line, M. Takinoue (Tokyo Institute of Technology) for productive discussions, A. Y. Hsiao (The Univ. of Tokyo) for useful comments on the manuscript, Y. J. Heo and S. Sugimoto (The Univ. of Tokyo) for useful suggestions regarding cell fibre transplantation, I. Obataya and N. Saito (JPK Instruments) for the measurement of hydrogel stiffness by atomic force microscopy, F. Ishidate for CLSM observation on NSC fibres, H. Teramae for physiological analyses of the paraffin sections and T. Hattori (Nippi) for fruitful advice on the material properties of collagen. We also thank M. Kiyosawa for the maintenance of diabetic mice and the measurement of their blood glucose concentrations, H. Aoyagi and M. Ishizaka for assistance in pancreatic islet isolation, M. Onuki for parylene deposition and A. Sato for drawing 3D computer graphics. This work was partly supported by the Takeuchi Biohybrid Innovation Project, Exploratory Research for Advanced Technology (ERATO), Japan Science and Technology (JST), Japan.

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H.O. and S.T. conceived the design of the study. H.O. fabricated cell fibres. T.O. and H.O. designed and conducted transplantation experiments of cell fibres to diabetic mice. A.I. contributed to the protein secretion analyses of cell fibres. M.K-N. and H.O. performed the Ca2+ imaging of the cortical cell fibre and the differentiation induction of the NSC fibre. R.G. and H.O. performed the knitting, reeling and weaving processes. H.O. and D.K. developed the double-coaxial microfluidic device and measured the mechanical strength of the cell fibres. K.S. performed the RT–PCR analysis and laser Raman scattering spectroscopy. S.M. examined the temporal changes of ECM proteins and the cell-to-cell contacts in the cell fibres. S.I. analysed Fourier-transform infrared spectra of the hydrogel materials. K.K-S. performed the folding of the cell fabric with H.O. Y.T.M. provided considerable advice on the initial direction of the research. Y.S. contributed to the development of the ECM-protein/Ca-alginate core–shell fibres. H.O., T.O. and S.T. wrote the paper. All authors discussed the results and commented on the manuscript.

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Correspondence to Shoji Takeuchi.

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The authors declare no competing financial interests.

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Onoe, H., Okitsu, T., Itou, A. et al. Metre-long cell-laden microfibres exhibit tissue morphologies and functions. Nature Mater 12, 584–590 (2013). https://doi.org/10.1038/nmat3606

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