Objective: To explore the interventional effect of platelet lysate (PL) on osteogenic differentiation of BMSCs by induction in rats in vitro., Methods: Twenty-four clean-grade adult Wistar rats, weighing from 250 g to 300 g, male or female, were included in this study. PL was obtained through three times of centrifugation and repeated freeze-thaw for the blood aspirated from cardiac cavities in 16 Wistar rats. ELISA assay was conducted to detect the concentration of growth factors PDGF, TGF-beta1, IGF-1 and VEGF in PL. The BMSCs harvested by flushing femurs of 8 adult Wistar rats were isolated, cultivated and expanded in vitro. The cells at the 4 passage were performed for osteogenic differentiation by induction in three groups of A (5% PL of final concentration in basic induction medium), B (1% PL of final concentration in basic induction medium), and C (no presence of PL in basic induction medium as a control). The morphological changes of the cells were dynamically observed with inverted phase contrast microscope during the whole period. At different time-points, ALP staining (7 days) and ALP/TP (2, 8, 12 days) of the cells were detected to evaluate ALP activity, and the mineral formation in extracellular matrix was examined with Alizarin red staining which provided quantitative analysis of mineral deposits., Results: ELISA assay showed that the content of PDGF, TGF-beta1, IGF-1 and VEGF in PL reached (300 +/- 30), (140 +/- 25), (80 +/- 35), (70 +/- 20) pg/mL, respectively. Morphological observation displayed BMSCs in group A or B gradually turned from spindle-shape to square- or polygon-shape as the morphological type of osteoblast-like cells at 7 days. The cells in group A showed slower shape changes but higher proliferation than that in group B or C. Moreover, at the 20 days, the cells in group A still displayed dense growth and produced obviously decreased amount of mineral deposits in ECM when compared with group B or C. At the 7 days, the cells of group A showed smaller amount of granules positive for ALP staining in cytoplasm when compared with groups B and C, and displayed marked reduction in ALP activity assay at the 2, 8, and 10 days compared with that of groups B and C (P < 0.05). At the 20 days, Alizarin red staining showed the number of mineral deposits in groups A, B and C were 7.67 +/- 1.10, 12.87 +/- 0.81 and 15.59 +/- 0.25, respectively, while the area of mineral deposits were (161 778.70 +/- 44 550.80), (337 349.70 +/- 56 083.24), and (415 921.70 +/- 71 725.39) pixels, respectively. The number of mineral deposits and the area of mineral deposits in group A were smaller than those in groups B and C (P < 0.05). But there was no statistically significant difference between groups B and C (P > 0.05)., Conclusion: PL is a kind of system carrying various growth factors. Exposure of PL inhibits both ALP activity and mineral formation of BMCs in a dose-dependent way under the osteogenic induction environment.