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Composable microfluidic spinning platforms for facile production of biomimetic perfusable hydrogel microtubes

Nature Protocols volume 16, pages 937–964 (2021)Cite this article

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Abstract

Microtissues with specific structures and integrated vessels play a key role in maintaining organ functions. To recapitulate the in vivo environment for tissue engineering and organ-on-a-chip purposes, it is essential to develop perfusable biomimetic microscaffolds. We developed facile all-aqueous microfluidic approaches for producing perfusable hydrogel microtubes with diverse biomimetic sizes and shapes. Here, we provide a detailed protocol describing the construction of the microtube spinning platforms, the assembly of microfluidic devices, and the fabrication and characterization of various perfusable hydrogel microtubes. The hydrogel microtubes can be continuously generated from microfluidic devices due to the crosslinking of alginate by calcium in the coaxial flows and collecting bath. Owing to the mild all-aqueous spinning process, cells can be loaded into the alginate prepolymer for microtube spinning, which enables the direct production of cell-laden hydrogel microtubes. By manipulating the fluid dynamics at the microscale, the composable microfluidic devices and platforms can be used for the facile generation of six types of biomimetic perfusable microtubes. The microfluidic platforms and devices can be set up within 3 h from commonly available and inexpensive materials. After 10–20 min required to adjust the platform and fluids, perfusable hydrogel microtubes can be generated continuously. We describe how to characterize the microtubes using scanning electron or confocal microscopy. As an example application, we describe how the microtubes can be used for the preparation of a vascular lumen and how to perform barrier permeability tests of the vascular lumen.

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Fig. 1: Schematic illustration of the microfluidic platform and the generated microtubes.
Fig. 2: Schematic representation of the assembly of the microfluidic devices.
Fig. 3: Spinning of microtubes with a straight or helical channel.
Fig. 4: Spinning of microtubes with double helical, double folded, straight-folded, or double straight channels.
Fig. 5: Spinning of microtubes with gas cavities and a straight channel.
Fig. 6: Spinning of microtubes with various knots.
Fig. 7: Representative images of the generated hydrogel microtubes.
Fig. 8: Representative images of cell-laden microtubes.
Fig. 9: Applications of the hydrogel microtubes.

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Acknowledgements

This work was financially supported by the National Natural Science Foundation of China (no. 81872835, 21621003), the Innovation Zone Project (18-163-12-ZT-003-077-01), and Health Major Project (BWS17J028, AWS16J018).

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Authors and Affiliations

  1. MOE Key Laboratory of Bioorganic Phosphorus Chemistry & Chemical Biology, Beijing Key Lab of Microanalytical Methods and Instrumentation, Department of Chemistry, Center for Synthetic and Systems Biology, Tsinghua University, Beijing, China

    Ruoxiao Xie, Zhe Liang, Yongjian Ai, Wenchen Zheng, Jialiang Xiong, Peidi Xu, Yupeng Liu, Mingyu Ding & Qionglin Liang

  2. China Astronaut Research and Training Center, Beijing, China

    Jianyi Gao & Jiaping Wang

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  1. Ruoxiao Xie
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Contributions

Q.L, R.X., Z.L., P.X., and Y.L. designed and performed the experiment; R.X. prepared the figures of the protocol; R.X. and Q.L wrote the protocol; Y.A, W.Z., J.X., M.D., J.G., and J.W. contributed to the discussion of the protocol.

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Correspondence to Qionglin Liang.

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Peer review Information Nature Protocols thanks Nicole Hashemi, Munenori Numata, and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Related links

Key references using this protocol:

Xu, P. et al. Adv. Mater. 29, 1701664 (2017): https://onlinelibrary.wiley.com/doi/abs/10.1002/adma.201701664

Xie, R. et al. Adv. Mater. 30, 1705082 (2018): https://onlinelibrary.wiley.com/doi/abs/10.1002/adma.201705082

Liu, Y. et al. RSC Adv. 8, 23475 (2018): https://pubs.rsc.org/en/content/articlelanding/ra/2018/c8ra04192j

Xie, R. et al. ACS Cent. Sci. 6, 903 (2020): https://pubs.acs.org/doi/abs/10.1021/acscentsci.9b01097

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Supplementary Data 1

AutoCAD file of the master mold for the custom-designed 96-well base plate.

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Xie, R., Liang, Z., Ai, Y. et al. Composable microfluidic spinning platforms for facile production of biomimetic perfusable hydrogel microtubes. Nat Protoc 16, 937–964 (2021). https://doi.org/10.1038/s41596-020-00442-9

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