Human mesenchymal stem cells can differentiate into various cell types and are useful for applications in regenerative medicine. Previous studies indicated that dental pulp exfoliated from deciduous teeth is a valuable alternative for dental tissue engineering because it contains stem cells with a relatively high proliferation rate. For clinical application, it is necessary to rapidly obtain a sufficient number of cells in vitro and maintain their undifferentiated state; however, the abundance of stem cells in the dental pulp tissue is limited. Non-thermal atmospheric pressure plasma (NTAPP) has been applied in regenerative medicine because it activates cell proliferation. Here, we examined the effects of NTAPP to activate the proliferation of human deciduous dental pulp fibroblast-like cells (hDDPFs) in vitro. Compared with untreated cells, NTAPP increased cell proliferation by 1.3-fold, significantly upregulated well-known pluripotent genes for stemness (e.g., Oct4, Sox2, and Nanog), and activated the expression of stem cell-specific surface markers (e.g., CD105). Overall, NTAPP activated the proliferation of various mesodermal-derived human adult stem cells while maintaining their pluripotency and stemness. In conclusion, NTAPP is a potential tool to expand the population of various adult stem cells in vitro for medical applications.