The isolation method for dental pulp stem cells (DPSCs) is still unclear to obtain a conducive environment for DPSCs to
proliferate. Enzymatic digestion and outgrowth method are two commonly used methods for DPSCs isolation but are not
well characterized in mice DPSCs. This study aimed to compare these isolation methods and differentiation potential
of mice DPSCs into bone cells. Dental pulp was extracted from mice’s incisors and subjected to isolation either by
collagenase 1A or culture of pulp tissue in complete alpha-Modified Eagle Medium (αMEM). Both cells isolated were
cultured until passage 4 and subjected to in vitro proliferation and differentiation analysis. Both cells exhibited fibroblastliked
morphology, but cells isolated by enzyme digestion proliferate faster compare to outgrowth method. After 21 days
of osteoblast differentiation, DPSCs isolated from enzyme digestion method showed alkaline phosphatase (ALP) activity
slightly different as compared to outgrowth method. In conclusion, there is a significant difference between the cells
isolated from enzyme digestion compare to outgrowth method with regard to proliferation and osteoblast differentiation.
Thus, it is preferable to isolate by enzyme digestion as it is faster and consistent compared to outgrowth method.