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Design of capillary microfluidics for spinning cell-laden microfibers

Nature Protocols volume 13, pages 2557–2579 (2018)Cite this article

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Abstract

This protocol describes the design of capillary microfluidics for spinning bioactive (cell-laden) microfibers for three-dimensional (3D) cell culture and tissue-engineering applications. We describe the assembly of three types of microfluidic systems: (i) simple injection capillary microfluidics for the spinning of uniform microfibers; (ii) hierarchical injection capillary microfluidics for the spinning of core–shell or spindle-knot structured microfibers; and (iii) multi-barrel injection capillary microfluidics for the spinning of microfibers with multiple components. The diverse morphologies of these bioactive microfibers can be further assembled into higher-order structures that are similar to the hierarchical structures in tissues. Thus, by using different types of capillary microfluidic devices, diverse styles of microfibers with different bioactive encapsulation can be generated. These bioactive microfibers have potential applications in 3D cell culture, the mimicking of vascular structures, the creation of synthetic tissues, and so on. The whole protocol for device fabrication and microfiber spinning takes ~1 d.

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Fig. 1: Schematic showing types of cell-laden microfibers that can be spun from a capillary microfluidic platform.
Fig. 2: Schematic illustrations of the capillary microfluidics.
Fig. 3: Images of the capillary microfluidics assembly procedure.
Fig. 4: Schematic illustrations and cross-sectional microscopy images of the microfibers.
Fig. 5: Schematic illustrations, and fluorescent and cross-sectional microscopy images of the spindle-knotted microfibers.
Fig. 6: Cell-laden microfibers for 3D cell culture and tissue-construct creation.

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Acknowledgements

This work was supported by the National Key Research and Development Program of China (2017YFA0700404), the National Science Foundation of China (grants 51522302 and 21473029), the NSAF Foundation of China (grant U1530260), the Fundamental Research Funds for the Central Universities, the Scientific Research Foundation of Southeast University, and the Scientific Research Foundation of the Graduate School of Southeast University.

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  1. These authors contributed equally to this work: Yunru Yu, Luoran Shang

Authors and Affiliations

  1. State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing, China

    Yunru Yu, Luoran Shang, Jiahui Guo, Jie Wang & Yuanjin Zhao

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Contributions

Y.Z. conceived the idea and designed the experiment. Y.Y. carried out the experiments. Y.Y., L.S., and Y.Z. analyzed the data and wrote the paper. J.G. and J.W. contributed to scientific discussion of the article.

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Correspondence to Yuanjin Zhao.

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Key references using this protocol

1. Onoe, H. et al. Nat. Mater. 12, 584–590 (2013): https://www.nature.com/articles/nmat3606

2. Cheng, Y. et al. Adv. Mater. 26, 5184–5190 (2014): https://onlinelibrary.wiley.com/doi/full/10.1002/adma.201400798

3. Cheng, Y. et al. ACS Appl. Mater. Interfaces 8, 1080–1086 (2016): https://pubs.acs.org/doi/abs/10.1021/acsami.5b11445

4. Wang, J. et al. Sci. China Mater. 60, 857–865 (2017): https://link.springer.com/article/10.1007/s40843-017-9081-5

Integrated supplementary information

Supplementary Figure 1 Schematic illustration and digital photographs of the tapered capillaries.

Schematic and digital photographs of the tapered capillary with a (a) tapered tip and (b) spindle tip.

Supplementary Figure 2 Close-up images of the capillary assembly procedure.

(a) Immobilize the square capillary on the glass slide; (b) Assemble the injection and collection capillaries inside the square capillary with epoxy resin; (c) Insert the spindle injection capillary inside the origin injection channel with epoxy resin; (d) Cut the needles with small crevices at the bottom, and make their sizes match with that of the capillaries; (e) Coat the bottom margin of the needles with a thin layer of epoxy resin, and stand them onto the junctions of the capillaries. The upper of the (a-c, e) images are the corresponding scheme illustrations.

Supplementary Figure 3 Images of the simple microfluidic device, the spinning process and the produced simple microfiber.

(a) Digital photograph of the simple microfluidic device; (b) The microstructure of the device during the simple microfiber spinning; (c) Digital image of the continuous simple microfiber in a vessel; (d) Microscopic image of the generated cylindrical microfiber. Adapted with permission from Cheng et al. Bioinspired multicompartmental microfibers from microfluidics. Adv. Mater. 26, 5184–5190 (copyright 2014 Wiley) (https://onlinelibrary.wiley.com/doi/abs/10.1002/adma.201400798). The scale bar is 200 μm.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–3

Reporting Summary

Supplementary Video 1

Creating tapered capillaries

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Yu, Y., Shang, L., Guo, J. et al. Design of capillary microfluidics for spinning cell-laden microfibers. Nat Protoc 13, 2557–2579 (2018). https://doi.org/10.1038/s41596-018-0051-4

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