2.5D tubulogenesis. Canis lupus familiaris

Hepatocyte growth factor-induced three-dimensional tubulogenesis (3D) is a simple and highly controllable system for studying epithelial tubule initiation and maintenance. However, due to its limited efficiency and asynchronous development, isolating genes associated with specific morphological changes observed during tubule formation has been unfeasible. Here we report a significantly enhanced in vitro culture method called 2.5-dimensional tubulogenesis (2.5D). Detailed image analysis of 2.5D has revealed morphologically distinct stages -- monolayer, extension, and tubule -- and finds that these stages display in a highly synchronized manner. Using time-course transcriptional profiling, we collected genes whose expression changes specifically associate with extension and tubule stage. Overall design: Time-course microarrays to identify genes whose temporal regulation is associated with extension and tubule stages. Four biological replicates for each stage (monolayer, extension, and tubule).

肝细胞生长因子诱导的三维管形成（3D）是一套用于研究上皮小管起始与维持过程的简易且高度可控的实验体系。然而，由于该体系效率有限且发育不同步，难以分离出与小管形成过程中特定形态变化相关的基因。本研究报道一种经显著优化的体外培养方法，命名为2.5维管形成（2.5D）。通过对2.5D体系的详细图像分析，我们发现其具有三个形态学特征明确的阶段：单层期、延展期与小管期，且各阶段呈现高度同步的发育进程。本研究通过时序转录谱分析，筛选出了与延展期和小管期特异性表达变化相关的基因。整体实验设计：采用时序微阵列芯片分析，以鉴定出时序调控模式与延展期和小管期相关的基因；每个阶段（单层期、延展期与小管期）均设置4个生物学重复。