Mini chamber system for long-term maintenance and observation of cultured cells

We constructed a mini chamber system that was able to maintain cell culture on a microscope for long periods. It is a modified closed system with medium perfusion and CO 2 circulation. The closed CO 2 circulation and ample air inside the chamber distinguish it from other closed systems. Using different cell lines, the system was shown to be able to support long-term, time-lapse recording. After 229 hours of time-lapse recording, A2058 cells (a melanoma cell line) became overconfluent but still multiplied. Many CAD cells (a murine neuron-like cell line) still moved their cell bodies and kept their neurite-like processes after 28 days of recording. The entire healing process of a scratch-wounded T24 (a bladder cancer cell line) monolayer can be monitored. Such a modified closed system should find many applications in developmental biology, cell biology, and cancer biology where long-term, time-lapse recording is required or when the health of cells is important.