The Effect of Different Concentrations of L-Ascorbic Acid on the Generation of Adipose-Derived Stem Cell Sheets

Adipose-derived stem cells (ADSCs), which are capable of self-renewal and multi-lineage differentiation, have most often been used in cell sheet engineering. L-ascorbic acid (LAA) plays an important role in the fabrication of the ADSCs cell sheet, but the optimal concentration of LAA to generate the ADSCs cell sheet and to regulate the secretion of extracellular matrix (ECM) components has not been well-studied. By supplementing different concentrations of LAA in the culture medium, we explored the effect of various LAA concentrations on adipose-derived stem cell sheet formation and detected the expression of type I collagen and fibronectin at both the gene and protein levels, in addition to observing the speed and thickness of cell sheet formation. After two weeks of cultivation, we demonstrated that LAA in the concentration range of 0-100 mu g/mL had no significant influence on the proliferation of ADSCs, but it could significantly enhance ADSCs cell sheet formation with abundant extracellular matrix deposition in a dose-dependent manner. Our results also showed that type I collagen was the principal component of the cell sheet induced by LAA stimulation, and the thickness could reach approximately 100 mu m at a concentration of 100 mu g/mL. We concluded that the optimal dose of LAA can provide a convenient and efficient approach to promote cell sheet formation for ASDCs mainly by increasing the secretion of type I collagen, and this information could be useful for researchers to optimize cell sheet characteristics for tissue engineering experiments.