Aim. This study aimed to investigate the potential of Liu’s Zhenggudan No. 2 Formula (LZF2) in inducing osteogenic differentiation of human umbilical cord blood mesenchymal stem cells (hUCB-MSCs) and treating osteoporosis (OP), thereby providing new methods and ideas for the treatment of OP by traditional Chinese medicine. Methods. Forty sample rats were equally divided into five groups: high-concentration LZF2, low-concentration LZF2, the Eucommia ulmoides (EU) group, the classical osteogenesis induction (COI) group, and the blank control group. Eight rats in each group were routinely housed for 7 days. Subsequently, to induce hUCB-MSCs, drug-containing serum was extracted from the abdominal aorta of rats to prepare the osteogenic induction solution. In addition, alkaline phosphatase (ALP) activity and osteocalcin (OCN) content assays, and alizarin red staining were performed on days 3, 6, 9, and 12 after culture. Results. After induction of hUCB-MSCs, ALP activity and OCN content increased significantly in the high-concentration LZF2 group. Alizarin red staining also depicted numerous orange-red calcified nodules in rats in the high-concentration LZF2 group. Conclusion. High concentration of LZF2 can facilitate the differentiation of hUCB-MSCs to promote osteogenesis.