(278h) 3-D Spheroid Model of Human Adipose-Derived Stem Cells for Superior Adipogenic Differentiation

Development of in vitro models of adipogenesis is critical to find therapeutics for obesity and related comorbidities. Here we present the 3-D spheroid model using human adipose-derived stem cells (hASCs) and their subsequent adipogenic differentiation. Our work describes construction of a 3-D spheroid culture system atop positively charged elastin-like polypeptide-polyethyleneimine (ELP-PEI) coating to study the differentiation and maintenance process of hASC adipocyte spheroids when subjected to fatty acid treatment. Measurements taken from spheroid micrographs indicated spheroid size increased during the maintenance phase, though the spheroids treated with linoleic acid or oleic acid had smaller size compared to those cultured in the control maintenance medium. Increased spheroid size correlated with increased triglyceride accumulation and the linoleic acid or oleic acid treatments stimulated triglyceride accumulation in hASC cultures. The CD36 expression, representing cell competency for consuming extracellular fatty acids, was consistently found in hASC 3-D spheroids. Overall, we observed that, the hASC spheroids undergo slower differentiation along the adipogenic lineage and attainment of physiological functionality compared to the 3T3-L1 mouse spheroids. Taken together, we have shown the feasibility of adipogenic differentiation of patient-derived hASC spheroids, which with further development, may help elucidate key features in the adipogenesis process.