Graduation Semester and Year

2010

Language

English

Document Type

Thesis

Degree Name

Master of Science in Biomedical Engineering

Department

Bioengineering

First Advisor

Young-Tae Kim

Abstract

It is very imperative to develop new in vitro platforms that can help in studying the complex interactions that occur between cell-Extracellular Matrix proteins and also during the process of cell migration in vivo. Such tools would facilitate in better understanding of the mechanisms that are employed by the cells during growth and migration. There are two devices that are presented in this study; 1) Multi-biomolecule coated lane device for unbiased cell preferential migration and 2) Novel Microchannel device for real time monitoring of tumor cell migration.The Multi-biomolecule coated lane device is a basic in vitro platform that helps in studying cell-biomolecules interaction using Microfluidics. This provides an unbiased cell-ECM interaction with up to 20 biomolecules as compared to the commercially available techniques that restrict to 2 or less at the same time. The device is fabricated using PDMS consisting of multiple protein lanes and a cell seeding area separated by PDMS barrier. Different ECM proteins (fibronectin, laminin, collagen type 1, vitronectin, bovine serum albumin, Aggrecan) were seeded into the lanes and were allowed to be absorbed on the substrate in cell culture incubator. Unbound proteins were washed and mammalian neuronal cells and humanvGlioblastoma Multiforme cells were seeded in the cell seeding area. It was seen that the cortical neurons showed a growth towards Laminin, fibronectin, collagen type 1, vitronectin and BSA but Aggrecan inhibited the growth of axons. DRG neurons and Schwann cells showed a robust growth towards the lanes containing collagen type 1, fibronectin and Laminin. The hGBM cells migrated and showed the maximum migration on Laminin and migration was also observed in lanes containing collagen type1, fibronectin and BSA. This platform presented here introduces a new scientific technique to study the interaction if different cells in interest towards proteins and other biomolecules to throw light on the complex interactions occurring in vivo.Novel Microchannel device for real time monitoring of tumor cell migration is a microfluidic device consisting of two different microchannel patterns; one with the channels tapering towards the distal end reservoir (from 20 μm, 15 μm, 10 μm, 8 μm to 5 μm) for studying single cell migration with respect to space. The other device design consists of adjacent lanes with different dimensions (20 μm, 15 μm, 10 μm, 8 μm and 5 μm). The device substrate was coated with laminin followed by seeding of primary human Glioblastoma Multiforme cells. Different time point images were taken during the course of cell migration through the channels and quantified for rate of migration. There was a significant difference in the rate of migration between the 5 um lane and 15 um. This serves as a new platform to understand the mechanisms involved in cancer migration with respect to availability of space and this would contribute in betterment of current treatment procedures.

Disciplines

Biomedical Engineering and Bioengineering | Engineering

Comments

Degree granted by The University of Texas at Arlington

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