The invention provides an umbilical cord Wharton's jelly mesenchymal stem cell osteogenic directional differentiation method and relates to the fields of stem cells and regenerative medicines. The method comprises the following steps of tissue digestion, primary culture, subculture, induction culture, etc.; in the tissue digestion step, tissue digestion liquid is used for tissue digestion, whereinthe tissue digestion liquid comprises collagenase I, DNA enzyme and Tryple; and in the induction culture step, an osteoblast differentiation induction culture medium is used for the induction culture, and the osteoblast differentiation induction culture medium comprises an alpha-MEM base bottom solution, a serum substitute, dexamethasone, beta-glycerophosphate, antibiotics, cytokine IL-beta, ascorbic acid and isoflavone. The method can obviously improve induction differentiation efficiency of umbilical cord mesenchymal stem cell osteoblasts.