The present invention discloses a polyurethane microsphere with diameter around 150 μm-270 μm, as well as the preparation method and use of it. The polyurethane which is prepared by this method has good biocompatibility, and it can be used as microcarrier to enhance cell proliferation. Meanwhile, the polyurethane microsphere is also injectable and enables to be used in tissue repair, evidently showing a well clinical application prospect.