Nahmias Yaakov, Odde David J
Center for Engineering in Medicine, Massachusetts General Hospital, Harvard Medical School, 51 Blossom Street, Boston, Massachusetts 02114, USA.
Nat Protoc. 2006;1(5):2288-96. doi: 10.1038/nprot.2006.386.
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.
在此，我们描述了一种简单的方案，用于设计和构建一种激光引导直接写入（LGDW）系统，该系统能够在体外以微米分辨率对肝窦样结构的自组装进行微图案化。据我们所知，LGDW是唯一能够在任意基质上，包括如基质胶这样的软凝胶上，以微米精度“即时”对细胞进行图案化的技术。通过在基质胶上对内皮细胞进行微图案化，可以控制血管结构和相关肝组织的自组装。因此，LGDW特别适合在单细胞分辨率下研究组织结构和力学性能的作用，以及研究诸如肝形态发生、分化和血管生成等过程中异型细胞间相互作用的影响。执行该方案所需的总时间通常为7小时。
