Culturing Murine Adipose-Derived Stem Cells as Spheroids in 5-Azacytidine and Trichostatin A Alters Gene Expression

Stem cells are undifferentiated cells that have the capability to differentiate into one or more cell lineages. Adipose-derived stem cells (ADSCs) are multipotent, mesenchymal stem cells that are located within the microvasculature of adipose tissue. Although multipotent ADSCs can differentiate into several cell lineages, they cannot match the differentiation potential of pluripotent stem cells, such as ES and iPS cells. However, previous research in our lab shows that culturing murine ADSCs as three-dimensional spheroids can induce the expression of genes associated with pluripotency. We hypothesized that the combination of culturing ADSCs as three-dimensional spheroids and treatment with compounds that manipulate the epigenome can 1) upregulate the expression of several genes associated with enhanced potency and 2) improve the efficiency of myogenic differentiation by ADSCs. Our results support our hypothesis that culturing ADSCs as spheroids in combination with treatment with trichostatin A, a histone deacetylase inhibitor, and 5-azacytidine, an inhibitor of DNA methylation, all impact the expression of genes associated with ADSCs’ potency and/or myogenic potential. Future work includes identifying the combination of culture conditions that most efficiently enhances the myogenic potential of ADSCs. This can be tested by recellularizing porcine acellular muscle matrix scaffolds with these enhanced ADSCs in order to assess their myogenic potential. Maximizing the myogenic potential of ADSCs would allow ADSCs to serve as a plentiful source of myogenic cells for skeletal muscle tissue engineering and regenerative medicine applications