Date of Graduation

2014

Document Type

Thesis

Degree Type

MS

College

Statler College of Engineering and Mineral Resources

Department

Lane Department of Computer Science and Electrical Engineering

Committee Chair

Yuxin Liu

Committee Co-Chair

Bourlai Thirimachos

Committee Member

Rajanasakul Yon

Abstract

Tubulogenesis and Angiogenesis is an important process in the formation of new blood vessels, and play a vital role in many physiological and pathological processes, such as cancer metastasis. To study this process in the laboratory, versatile, reliable and affordable assays are essential. The objective of this thesis work was to develop a microfluidic three-dimensional (3D) gradient device, which embedded collagen within the device for generating chemical gradient through it, and investigated it as a useful research tool for angiogenesis study with endothelial cells forming branching tubes inside the collagen.;With the enormous growth of microfluidic technologies, the scope for creating more realistic in vitro cell angiogenesis assays that replicate many aspects of the true in vivo microenvironment has increased. Although few conventional assays like rabbit ear chamber assay and chick chorio allantoic membrane assay are available in market, they set their own limitations for further studies. For instance, high in cost, lack of precise gradient control and characterization, limitation in mimicking the micro environment etc. Here, in this thesis work we introduced a microfluidic 3D gradient device and angiogenesis study assay that serves as a versatile single platform for the study of angiogenesis which is crucial step in the study of vascular biology and it related diseases. Briefly, a uniform collagen layer, which served as an extra cellular matrix (ECM), gave a precise control of generating a chemical gradient over a long period of time and offered an excellent monitoring capability with response to observing endothelial cells' formation of tube structures. Human umbilical vein endothelial cells (HUVEC) were cultured inside the corresponding location of a micro channel for 4-5 days, and their responses to the quantified gradient of vascular endothelial growth factor (VEGF) were examined. Immunofluorescence staining and confocal imaging were made for detailed study on formed tubing. These results suggested that the microfluidic 3D gradient device can conveniently generate a stable chemical gradient and provide an easy way for the study of angiogenesis.

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