Single-cell cloning and expansion of human induced pluripotent stem cells by a microfluidic culture device

• We developed a microfluidic device to maintain cells in long-term cultures.
• The device was designed to recover cells for further analyses.
• The culture conditions of human induced pluripotent stem cells were improved.
• Clonality-validated cell lines were successfully established with the device.

The microenvironment of cells, which includes basement proteins, shear stress, and extracellular stimuli, should be taken into consideration when examining physiological cell behavior. Although microfluidic devices allow cellular responses to be analyzed with ease at the single-cell level, few have been designed to recover cells. We herein demonstrated that a newly developed microfluidic device helped to improve culture conditions and establish a clonality-validated human pluripotent stem cell line after tracing its growth at the single-cell level. The device will be a helpful tool for capturing various cell types in the human body that have not yet been established in vitro.