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Micropatterning of living cells by laser-guided direct writing: application to fabrication of hepatic–endothelial sinusoid-like structures

Nature Protocols volume 1, pages 2288–2296 (2006)Cite this article

Abstract

Here, we describe a simple protocol for the design and construction of a laser-guided direct writing (LGDW) system able to micropattern the self-assembly of liver sinusoid-like structures with micrometer resolution in vitro. To the best of our knowledge, LGDW is the only technique able to pattern cells “on the fly” with micrometer precision on arbitrary matrices, including soft gels such as Matrigel. By micropatterning endothelial cells on Matrigel, one can control the self-assembly of vascular structures and associated liver tissue. LGDW is therefore uniquely suited for studying the role of tissue architecture and mechanical properties at the single-cell resolution, and for studying the effects of heterotypic cell–cell interactions underlying processes such as liver morphogenesis, differentiation and angiogenesis. The total time required to carry out this protocol is typically 7 h.

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Figure 1: LGDW system.
Figure 2: Developing umbilical vein endothelial cell pattern imaged via brightfield microscopy.
Figure 3: Final pattern of umbilical vein endothelial cells written on a collagen-coated surface by LGDW, 24 h after guidance.
Figure 4: LGDW of vascular structures on Matrigel.
Figure 5: LGDW of liver sinusoid-like structures.
Figure 6: Hepatic–endothelial organoid formation.

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Authors and Affiliations

  1. Center for Engineering in Medicine, Massachusetts General Hospital, Harvard Medical School, 51 Blossom Street, Boston, 02114, Massachusetts, USA

    Yaakov Nahmias

  2. Department of Biomedical Engineering, University of Minnesota, 7-132 Nils Hasselmo Hall, 312 Church Street SE, Minneapolis, 55455, Minnesota, USA

    David J Odde

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  1. Yaakov Nahmias
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Correspondence to David J Odde.

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Nahmias, Y., Odde, D. Micropatterning of living cells by laser-guided direct writing: application to fabrication of hepatic–endothelial sinusoid-like structures. Nat Protoc 1, 2288–2296 (2006). https://doi.org/10.1038/nprot.2006.386

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