DNA origami is a newly proposed DNA self-assembly method in recent years. It can design specific DNA sequences and follow the principle of complementary base pairing to construct more complex nanostructures and nanopatterns. The recent discovery of tetrahedral DNA nanostructure (TDN) constructed by DNA origami has great application potential in the field of stem cells. In this study, we reviewed the structure and biological properties of TDN, and the applications of TDN in the field of stem cells.