Osteogenic differentiation is a process of bone marrow mesenchymal stem cells (BMSCs), periosteal stem cells (PSCs), adipose-derived stem cells (ADSCs), and other cell types differentiating into osteoblasts, which are bone-forming cells that synthesize and mineralize the extracellular matrix. The differentiation of osteoblasts is a complex process that is controlled by kinds of signaling pathways and orchestrated by various cells in the osteogenic microenvironment to maintain the balance in bone metabolism. Currently, the components to induce osteogenic differentiation include cytokines, hormones, biological materials, and physical stimulation. Understanding and standardizing the isolation and culture methods can provide a theoretical basis for the use of bone tissue engineering technology and accelerate the treatment process of bone diseases such as osteoporosis and bone fracture. Examples of methods to be found in this collection are animal models such as the femoral fracture model, tibial monocortical defect model, and cranial defect model, isolation of chondrocytes, vascular endothelial cells, and immune cells in the osteogenic microenvironment, approaches of stem cells co-culture, the induction and identification of osteogenic differentiation, etc.