Background: Microarray gene expression technology has become standard practice in systems biology to analyze transcriptomic changes at the cell, tissue, and organism level, enabling the reverse-engineering of condition-specific gene networks. Purpose: To better understand how transcriptional gene networks of biologically close cellular systems, differ, two specific networks were reverse-engineered from 2D and 3D human normal bronchial epithelial cell cultures. Methods: 100 gene expression profiles for both normal human bronchial epithelial (NHBE) cells and organotypically differentiated bronchial cells (AIR100 culture) were collected under normal culture conditions. Gene networks were reconstructed by correlating gene expression profiles at a false discovery rate (FDR, T-statistic and Benjamini-Hochberg correction).