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

## TL;DR

This paper introduces a new method to study how zebrafish vertebrae grow in a lab setting, mimicking natural growth patterns.

## Contribution

The study is the first to enable live analysis of fish vertebral growth from late larval stages using air-liquid interface culture.

## Key findings

- Air-liquid interface culture successfully reproduces the mineralization patterns of in vivo vertebral growth.
- The method allows observation of vertebral growth and related cell behavior that was previously difficult to study in live fish.

## Abstract

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.