Todhunter Michael E, Jee Noel Y, Hughes Alex J, Coyle Maxwell C, Cerchiari Alec, Farlow Justin, Garbe James C, LaBarge Mark A, Desai Tejal A, Gartner Zev J
Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, California, USA.
Tetrad Graduate Program, University of California, San Francisco, San Francisco, California, USA.
Nat Methods. 2015 Oct;12(10):975-81. doi: 10.1038/nmeth.3553. Epub 2015 Aug 31.
Reconstituting tissues from their cellular building blocks facilitates the modeling of morphogenesis, homeostasis and disease in vitro. Here we describe DNA-programmed assembly of cells (DPAC), a method to reconstitute the multicellular organization of organoid-like tissues having programmed size, shape, composition and spatial heterogeneity. DPAC uses dissociated cells that are chemically functionalized with degradable oligonucleotide 'Velcro', allowing rapid, specific and reversible cell adhesion to other surfaces coated with complementary DNA sequences. DNA-patterned substrates function as removable and adhesive templates, and layer-by-layer DNA-programmed assembly builds arrays of tissues into the third dimension above the template. DNase releases completed arrays of organoid-like microtissues from the template concomitant with full embedding in a variety of extracellular matrix (ECM) gels. DPAC positions subpopulations of cells with single-cell spatial resolution and generates cultures several centimeters long. We used DPAC to explore the impact of ECM composition, heterotypic cell-cell interactions and patterns of signaling heterogeneity on collective cell behaviors.
从细胞构建单元重构组织有助于在体外对形态发生、体内平衡和疾病进行建模。在此，我们描述了细胞的DNA编程组装（DPAC），这是一种重构具有程序化大小、形状、组成和空间异质性的类器官样组织的多细胞组织的方法。DPAC使用经可降解寡核苷酸“维可牢”化学功能化的解离细胞，使细胞能够快速、特异性且可逆地粘附到其他涂有互补DNA序列的表面。DNA图案化底物充当可移除且有粘性的模板，逐层DNA编程组装将组织阵列构建到模板上方的三维空间中。DNA酶从模板中释放出完整的类器官样微组织阵列，同时将其完全包埋在各种细胞外基质（ECM）凝胶中。DPAC以单细胞空间分辨率定位细胞亚群，并生成数厘米长的培养物。我们使用DPAC来探究ECM组成、异型细胞间相互作用以及信号异质性模式对集体细胞行为的影响。
