Hyperglycemia impairs skeletogenesis from embryonic stem cells by affecting osteoblast and osteoclast differentiation

High maternal blood glucose levels caused by diabetes mellitus can irreversibly lead to maldevelopment of the growing fetus with specific effects on the skeleton. To date, it remains controversial at which stage embryonic development is affected. Specifically during embryonic bone development, it is unclear whether diminished bone mineral density is caused by reduced osteoblast or rather enhanced osteoclast function. Therefore, the aim of this study was to characterize the growth as well as the skeletal differentiation capability of pluripotent embryonic stem cells (ESCs), which may serve as an in vitro model for all stages of embryonic development, when cultured in diabetic levels of D-glucose (4.5 g/L) versus physiological levels (1.0 g/L). Results showed that cells cultivated in physiological glucose gave rise to a higher number of colonies with an undifferentiated character as compared to cells grown in diabetic glucose concentrations. In contrast, these cultures were characterized by slightly decreased expression of proteins associated with the stem cell state. Furthermore, differentiation of ESCs into osteoblasts and osteoclasts was favored in physiological glucose concentrations, demonstrated by an increased matrix calcification, enhanced expression of cell-type-specific mRNAs, as well as activity of the cell-type-specific enzymes, alkaline, and tartrate resistant acidic phosphatase. In fact, this pattern was noted in murine as well as in primate ESCs. Our study suggests that an interplay between both the osteoblast and the osteoclast lineage is needed for proper skeletal development to occur, which seems impaired in hyperglycemic conditions.