Development of 3D Microfluidic Living Cell Arrays For Anti-Cancer Drug Screening in Mimicked Tumor Microenvironment
Item
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Title
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Development of 3D Microfluidic Living Cell Arrays For Anti-Cancer Drug Screening in Mimicked Tumor Microenvironment
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Identifier
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d_2009_2013:6c30de21ae36:11718
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identifier
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12321
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Creator
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Dereli-Korkut, Zeynep,
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Contributor
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Sihong Wang
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Date
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2013
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Language
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English
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Publisher
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City University of New York.
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Subject
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Biomedical engineering | Pharmaceutical sciences | 3D Microfluidic cell array | apoptosis | caspase-3 | caspase-8 | reporter cell line | tumor microenvironment
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Abstract
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The promise of improved cancer therapy has been one of the driving forces for cell death research over the past decade. There is growing evidence that many of the molecular and cellular changes that occur in cancer development diminish the ability of cells to undergo apoptosis and thus cause drug resistance. Therefore, developing and screening novel anticancer drugs that target apoptosis pathways have received increasing attention in the past few years. However, identification of novel compounds and drug targets involved in apoptosis regulation is still a major roadblock to anticancer drug development due to the lack of a high throughput apoptotic screening system which can systematically measure dynamic expression of multiple proteins and genes as well as enzyme activities in real time in intact cells from multiple stimuli. To systematically profile apoptotic signaling pathways, we proposed to dynamically monitor the behavior of several genes and caspases in a real-time by apoptosis reporter cell lines. Meanwhile, a microfluidic device was developed to enable 3 dimensional co-culture of tumor cells with microvascular endothelial cells to better mimic the in vivo microenvironment in vitro.;In this thesis, the development of apoptosis reporter cell lines was focused on three molecules in apoptosis pathways, caspase 8, BcL-xl and NFkappaB. The tumor microenvironment has a critical impact on anticancer therapies. Here, the development of a three dimensional Microfluidic Living Cell Array (3D MLCA) to model the tumor microenvironment was reported. The 3D MLCA consists of three layers microfabricated using polydimethylsiloxane (PDMS). The bottom layer of the device includes cell culture chambers, the middle is a permeable filter layer and the upper layer includes microchannels for fluid flow. Caspase 3 activities of multiple anti-cancer reagents were used to evaluate the device. The transport and exchange of nutrients and waste products between long-term cultured human Microvascular Endothelial Cells (HMVEC) and non-small cell lung cancer cells (PC9) embedded in peptide hydrogel (Puramatrix) were successfully maintained using our 3D microfluidic cell arrays. Direct visualization and quantitative analysis of caspase 3 activities in co-cultured PC9 cells and endothelial cells exposed to four anti-cancer drugs were a confirmation of the versatility of our system for biological process assessments. We compared drug test results of PC9 and endothelial cell co-culture in a 3D MLCA with conventional tissue culture plates. This 3D MLCA provides a proof of concept for mimicking tumor microenvironment at a high throughput level.;Combining apoptosis reporter cell lines with 3D midrofluidic cell arrays will enable more accurate anti-cancer drug screening at a high throughput level in the mimicked 3D tumor microenvironment. Thus, the therapeutic prediction provided from the drug screening would offer better outcomes in a clinical setting.
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Type
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dissertation
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Source
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2009_2013.csv
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degree
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Ph.D.
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Program
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Engineering
