The invention relates to a method of acquiring hematopoietic stem cells (HSCs) by means of three dimensional inducing system with high efficiency—which makes use of three-dimensional cell culture matrix or cell culture scaffolds, such as three-dimensional cell culture system which is made from hydrogel, seaweed and other materials, and / or associating stromal-cell including bone marrow cell, mouse bone marrow cell line OP9, OP9DL1 and so on, and / or associating multiple factors including mesoderm inducible factor, hematopoietic growth factor etc which can induce pluripotent stem cells to differentiate into hematopoietic stem cells. The invention sets up a new method of acquiring hemopoietic stem cells, establishing a system that induces pluripotent stem cells to differentiate into hemopoietic stem cells with high efficiency by means of utilizing dimensional inducing system and / or associating stromal-cell including bone marrow cell and / or multiple factors for the first time, which provide a theoretical basis and technology platform for acquiring clinical available hematopoietic stem cells, and expand additional methodologies and ideas in fields of disease mechanism exploring, drug screening etc for hematopoietic cells from pluripotent stem cells.