Department of Chemical and Biological Engineering, Princeton University, Princeton, NJ, USA.
Department of Molecular Biology, Princeton University, Princeton, NJ, USA.
Methods Mol Biol. 2024;2805:113-124. doi: 10.1007/978-1-0716-3854-5_7.
The extracellular matrix (ECM) provides dynamic structural and molecular signals that affect the form and function of developing tissues. In order to parse how the individual features of the ECM impact cell- and tissue-level behavior during development, engineered culture models should reproduce key structural and molecular features of native ECM. Here, we describe a protocol for bioprinting epithelial cell aggregates embedded within a collagen-Matrigel ink in order to study the dynamic interplay between epithelial tissues and aligned networks of type I collagen fibers. Collagen fiber alignment and geometry can be spatially controlled by modulating the printing speed, nozzle geometry, surface chemistry, and degree of molecular crowding in the printing ink. We provide detailed procedures for generating epithelial cell aggregates, microextrusion printing collagen-Matrigel bioinks, culturing the three-dimensional (3D)-printed tissues, and imaging 3D-printed collagen-Matrigel constructs.
细胞外基质 (ECM) 提供动态的结构和分子信号，影响发育组织的形态和功能。为了分析 ECM 的各个特征如何在发育过程中影响细胞和组织水平的行为，工程化的培养模型应再现天然 ECM 的关键结构和分子特征。在这里，我们描述了一种在胶原-基质胶墨水中生物打印上皮细胞聚集体的方案，以便研究上皮组织与排列整齐的 I 型胶原纤维网络之间的动态相互作用。通过调节打印速度、喷嘴几何形状、表面化学性质和打印墨水中的分子拥挤程度，可以对胶原纤维的排列和几何形状进行空间控制。我们提供了详细的程序来生成上皮细胞聚集体、微挤出打印胶原-基质胶生物墨水、培养三维 (3D) 打印组织以及对 3D 打印胶原-基质胶结构进行成像。
