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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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Three-Dimensional Co-Culture Systems for Vascularization of Cardiac Tissue
Title
Three-Dimensional Co-Culture Systems for Vascularization of Cardiac Tissue
Creator
Rodriguez Arias, Jessica A.
Date
2023
Description
Myocardial Infarction (MI) is the partial or complete blockage of blood flow to the myocardial tissue resulting in damage and therefore loss...
Show moreMyocardial Infarction (MI) is the partial or complete blockage of blood flow to the myocardial tissue resulting in damage and therefore loss of heart function. In the U.S. every 40 seconds, someone will suffer from MI and the only available treatment is medication to treat the symptoms of heart function loss, but do not treat the underlying cause. Some attempts to treat the underlying cause have arisen in the last decades including cell-based therapies or tissue engineering therapies such as spheroid-based cardiac patches that have shown to be promising. Improvement in the mechanical properties to create suturable engineered tissues remain to be improved for ease of implantation purposes. Cell-laden hydrogel scaffolds can provide improved mechanical properties compared to biomaterial free cell-based therapies but need to allow for vascularization of the engineered tissue. Thus, the goal of this thesis is to provide preliminary studies for the use of a cell adhesive, proteolytically degradable PEG hydrogel scaffold that eventually would be used as an invitro model to evaluate engineered tissue vascularization for cardiac tissue engineering. To construct this model, important cell spheroid parameters on vascular invasion in 3D culture were investigated including the total number of cells/spheroid, the supporting cell for endothelial cells. In order to scale-up scaffolds to size of clinically relevant dimensions, a multilayered hydrogel construct visible light free-radical polymerization approach encapsulating vascular spheroids in multiple layers was also investigated. Results indicate that a total cell number of 5000 cells/spheroid aggregate were feasible due to cell sourcing. In addition, co-cultures of endothelial and mesenchymal stem cells led to maximized vascular invasion of the spheroids compared to fibroblast/endothelial co-culture and endothelial monoculture of spheroids in the hydrogel. Finally, the extent of vascularization of spheroids in each layer of the multilayered hydrogel constructs varied due to the observed differences in mechanical properties and swelling ratio of each layer due to incomplete polymerization of layers. This study demonstrated the importance of support cells and hydrogel mechanical properties in promoting vascularization of spheroid which serves as basis for building cell-laden hydrogel scaffolds for vascularization for cardiac tissues.
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