Adipose tissue models can be used for in vitro drug screening of therapeutics designed for the treatment of obesity or adipose tissue-related... Show moreAdipose tissue models can be used for in vitro drug screening of therapeutics designed for the treatment of obesity or adipose tissue-related diseases. This work aimed to engineer functional three-dimensional (3D) adipose microtissue models that could be incorporated within a microfluidic system. To support the on-chip 3D culture, a microfluidic device consisted of cell culture chambers flanked by two side channels was designed. The mold for the microfluidic device was manufactured using computer numeric control (CNC) micro-milling. Soft lithography with polydimethylsiloxane (PDMS) was used to construct the microchannels and chambers in the microfluidic device. A model was developed by the monoculture of adipocytes within the microfluidic device. Human adipose-derived stem cells (ADSCs) were differentiated toward adipocyte in the cell culture chambers and formed a 3D adipose microtissue. The effect of interstitial flow on the adipogenic differentiation of ADSCs was explored. Adipocytes showed decreased adiponectin secretion and increased lipolysis in response to increased interstitial shear stress. Meanwhile, multiple adipogenic genes were downregulated with the increase in shear stress.To engineer vascularized adipose tissue, a co-culture system with ADSCs, human umbilical vein endothelial cells (HUVECs) and normal human lung fibroblasts (NHLFs) was applied. Culture conditions (media, cell ratios, temporal conditions, etc.) for optimal differentiation of ADSCs and induction of network formation were identified. ADSCs were induced toward adipogenesis before mixed with HUVECs and NHLFs. The cell mixture was loaded into the microfluidic device and formed an adipose microtissue with a vessel network in a mixed culture media. An interconnected vascular network was established within 2 weeks and formed anastomoses with the side channels. Perfusion of fluorescent dextran confirmed the interconnections and lumen formation of the vascular network. Perfusion of fluorescently labeled fatty acid analog through vessels resulted in the accumulation of the fatty acid in adipocytes, confirming the functionality of the adipose microtissue. In conclusion, this work presented adipose tissue models within a microfluidic device that can potentially be utilized for on-chip drug screening, as well as provide insights into the engineering of complex tissues. Show less
