Abstract: Objective：Studies on the effect of cryopreservation on the biological characteristics of dentine and dental pulp cells in vitro. Methods：A total of 40 fresh teeth (the first premolar or second premolar) were collected and respectively experiment with dentine mechanical morphological analysis and dental pulp cells growth experiments. At the experiment of dentine mechanical morphological analysis, 20 fresh extracted human premolars were selected，prepared into dentin discs，and randomly divided into 2 groups (group A：fresh teeth group，group B：programmed cooling group), which were measured for compression strength and scanning electron microscope analysis. Another 20 teeth, using improved tissue block method of separation of dental pulp cells, the original generation of cryopreserved after recovery were observed before and after analysis of cryopreserved cells morphological and growth characteristics. Results：There were no obvious differences between groups with cryopreservation in growth characteristic of dentine and dental pulp cells. Conclusions：Tooth pulp tissue can keep good biological characteristics with cryopreservation.