Abstract: The present disclosure relates to a method for producing mesenchymal stem cells from pluripotent stem cells and mesenchymal stem cells prepared by the method. The present disclosure enables mesenchymal stem cells having superior proliferation rate while having superior intrinsic biological activity to be obtained with high yield by sequentially performing three-dimensional suspension culture using pluripotent stem cells, particularly induced pluripotent stem cells (iPSCs), as starting cells and adherent culture of cell aggregates formed therethrough. The mesenchymal stem cells produced by the method of the present disclosure can be usefully used in compositions for treating bone diseases, cartilage diseases or inflammatory and autoimmune diseases owing to high differentiation efficiency into bone and cartilage and superior anti-inflammatory activity.