Background : Though the treatment of atrophic nonunion is often challenging, bone union may occurs in some patients with atrophic nonunion who underwent indirect treatment at the docking site. We aimed at explaining the reasons why does this phenomenon appear? Methods : Five patients diagnosed with atrophic nonunion were enrolled in our study. Nonunion tissues were cut into strips and cultured immediately after being obtained. We got the adherent cells from atrophic nonunion tissues, and applied the flow cytometry to assess the expression of different cell-surface protein. The research in differentiation of the adherent cells was carried out under the induction of various lineage-specific factors, and Western-blot analysis was used to measure the differentiation-related protein expression. Results : We found that the adherent cells from atrophic nonunion tissues have the characteristics similar to bone marrow stromal cells. These traits demonstrated by the flow cytometry consist of positive expression of markers CD29 and CD44, but negative expression of CD34 and CD45. The adherent cells could differentiate into osteogenic, chondrogenic, and adipogenic cells under the different lineage-specific induction factors in vitro. Alkaline Phosphatase (ALP), Col Ⅱ, osteocalcin (OC), SOX9, lipoprotein lipase (LPL) and PPAR-γ2 were high expressed, as measured by Western-blotting. Conclusions : Mesenchymal stem cells obtaining from atrophic nonunion tissues have the potential of transforming into cartilage and bone-forming cells. Furthermore, we get conclusion that the atrophic nonunion tissues play an important role in the healing process.