The CaO-ZrO2-SiO2 (CZS) coating was prepared by plasma-spraying chemically synthesized CZS powder onto a Ti-6Al-4V substrate. This CZS coating has been demonstrated to have good bioactivity, high bonding strength with the substrate and a low degradation rate. However, the effect of CZS coating on the osseointegration of bone-implant is still unknown. In this study, human bone marrow-derived stromal cells (hBMSCs) were cultured on CZS coating in vitro, and cell behavior was investigated, with the classical hydroxyapatite (HA) coating as a control. Scanning electron microscopy (SEM) and immunofluorescence studies showed that the hBMSCs on the CZS coating spread well with organized cytoskeleton structure at 24 h following cell seeding. The MTT assay and the Alamar Blue assay indicated that CZS coating promoted the attachment and proliferation of hBMSCs. The results of alkaline phosphatase (ALP) activity test and the expression of osteogenic marker genes, such as ALP, collagen I (COLI), osteopontin (OPN), and osteocalcin (OCN), demonstrated that the osteoblastic differentiation of hBMSCs was enhanced more by CZS coating than by HA coating. These results suggest that CZS coating possess excellent biological properties and may have potential in biomedical applications.