Air-liquid interface culture for live analysis of zebrafish vertebral growth

Fish vertebrae change their shapes from simple cylinders to complex amphicoelous hourglass-like forms with lateral ridges as they grow. This growth process is difficult to observe in vivo due to the thickening of the muscles surrounding the vertebrae. In this study, we applied air-liquid interface culture to isolated zebrafish vertebrae to observe vertebral growth in vitro . The mineralization patterns of the cultured vertebrae were similar to those of in vivo growth, reproducing vertebral growth. This study is the first report to ensure live analysis of fish vertebral growth from late larval stages and the related cell behavior.