High-throughput formation and long-term culture of embryoid bodies using a microfluidic hanging drop chip

In this paper, we present a microfluidic chip-based method for high-throughput formation and long-term culture of embryoid bodies (EBs). The microfluidic chip is designed to be a stand-alone device whose operation can be performed by a manual pipette, making it easy to use in a regular biology laboratory setup. We show that uniform-sized mouse EBs can be formed after 1 day of culture in the microfluidic hanging drop chip. By exchanging the medium in the chip once every 2 days during culture, the EBs could be cultured on chip for up to 7 days. The immunostaining result of the normal germ layer formation of the EBs after the 7 days culture, showing the potential of our microfluidic chip to be a useful tool for long-term induction of EB differentiation.