Autogenous particulate bone grafts are being utilized in oral implantology for minor grafting procedures. This study aimed to investigate the influence of the bone-harvesting technique, donor age, and donor site on proliferation and differentiation of human primary osteoblast-like cells in the cell culture. Autogenous bone particles (20 samples) were harvested from the maxilla and mandible during surgery using two different protocols, and two types of particulate bone grafts were collected: bone chips and bone sludge. Bone samples were cultured in growth medium and, after 2 to 3 weeks, the cells that grew from bone grafts were cultured in the normal and osteogenic medium for 0, 4, 7, and 20 days. DNA, alkaline-phosphatase (ALP), calcium-content measurements, and Alizarin red/toluidine blue staining were performed. Data were analyzed by repeated-measures analysis of variance with Bonferroni test. The level of statistical significance was set at 5% (P < .05). Total DNA, ALP, and calcium content were significantly higher for the bone chip samples compared to the bone sludge samples. Total DNA and ALP content were significantly higher for the patients in age group 1 (≤ 60 years) compared to age group 2 (> 60 years) and was significantly higher for mandibular samples than maxillary samples on day 20. However, the calcium measurement showed no significant difference concerning donor age and donor site. Data analysis revealed that harvesting technique (bone chips vs bone sludge), donor age (≤ 60 years vs > 60 years), and donor site (maxilla vs mandible) influenced the osteogenic potential of the collected particulate bone graft. The bone chips were superior in terms of osteogenic efficacy and should be considered a suitable option for particulate bone graft collection.