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ENGINEERING HUMAN ADIPOSE TISSUE WITHIN A MICROFLUIDIC DEVICE
Title
ENGINEERING HUMAN ADIPOSE TISSUE WITHIN A MICROFLUIDIC DEVICE
Creator
Yang, Feipeng
Date
2019
Description
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.
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A MICROFLUIDIC INTESTINAL-MICROBIOTA PLATFORM TO STUDY DRUG METABOLISM
Title
A MICROFLUIDIC INTESTINAL-MICROBIOTA PLATFORM TO STUDY DRUG METABOLISM
Creator
Wang, Chengyao
Date
2020
Description
The intestine is the main site that orally administered drugs are primarily metabolized, absorbed, and distributed. The trillions of bacteria...
Show moreThe intestine is the main site that orally administered drugs are primarily metabolized, absorbed, and distributed. The trillions of bacteria that inhabit the intestine influence health and regulate important biochemical factors, such as the activity of enzymes pertinent to drug metabolism. However, this has not been systematically studied partly due to the challenges of recapitulating the unique and complex intestinal microenvironment that includes (1) the presence of mammalian and microbial cells and (2) a unique partitioned oxygenation proļ¬le across the lumen to the subepithelial mucosa from anaerobic to the richly vascularized oxygenated. This thesis reports the development of a microfluidic device in which is integrated a membrane synthesized from a key element of mucosal basal lamina, collagen, and precisely controlled partitioned oxygen environment. The device enabled excellent cell viability and long-term function. More importantly, it enabled the coculture of intestinal epithelial cells and aerobic and anaerobic bacteria in the partitioned oxygen environment. These experiments on one hand allowed the measurement of cellular oxygen consumption rate under perfusion, which could be used to study microbial regulation of oxidative metabolism in epithelial cells. On the other hand, the device allowed a systematic examination of the role of different gut bacteria strains on the regulation of factors that are important in drug metabolism, namely, transporters and phase I enzymes. Our studies highlighted the importance of direct communication between the intestinal cells and the gut bacteria with major findings being that species-specific differences exist in the regulation of drug metabolism. This work will be useful for (1) the discovery of novel regulators of drug metabolizing enzymes, (2) developing new pharmacokinetic models, and (3) advancing precision medicine models for patients.
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