Patients with type 2 diabetes mellitus experience a slower healing process and poor osteointegration, making it difficult for them to heal properly after a bone fracture. This study aims to compare the proliferation and differentiation of human mesenchymal stromal cells at different glucose concentrations, as well as with an advanced glycated end-product (AGE) concentration, to mimic a healthy, prediabetic, and diabetic environment in an in vitro model over several experiments. Each experiment was composed of treatment groups in either growth or osteogenic media, with varying levels of glucose concentration or an advanced glycated end-product concentration. The treatment groups were cultured in 24 well plates over 28 days with staining of FITC-maleimide, DAPI, or alkaline phosphatase conducted at varying time points. The plates were imaged, then analyzed in ImageJ and GraphPad Prism. The study supports that at 28 days in culture, the more glucose added to osteogenic media treatment groups, the lower the nuclear count. At 14 days the same is true of growth media treatment groups, though the trend does not persist until 28 days. It does not seem that cell surface area of osteogenic groups, and growth media treatment groups was affected by glucose level. At 14 days, the alkaline phosphatase expression was unaffected by glucose level. However, at the 28 day time point the higher the glucose level of osteogenic treatment groups, the less expression of alkaline phosphatase. The effect of the added AGE concentration on hMSC osteogenesis was inconclusive. Overall, this study enhanced understanding of the role that glucose and AGEs play in the bone healing process for diabetic patients, allowing for future improvements of biomaterials and engineered tissue.