Disclosed are a medium composition for culturing mesenchymal stem cells for the treatment of cancer which can inhibit proliferation of cancer cells, while maintaining differentiation capability and activity thereof, and a method for producing mesenchymal stem cells for the treatment of cancer using the composition. More particularly, disclosed are a medium composition containing vitamin C and aspirin for producing mesenchymal stem cells having improved inhibitory activity against proliferation of cancer cells, and a method for producing mesenchymal stem cells for the treatment of cancer using the composition.