Title Page
Contents
ABSTRACT 9
INTRODUCTION 10
Materials and Methods 12
2.1. Microfluidic chamber design and fabrication 12
2.2. Establishment of hypoxic condition within dynamic microfluidic chamber 14
2.3. Cell culture 15
2.4. Cell viability and cancer cell spheroid formation 16
2.5. Observation of NK cell migration under dynamic microfluidic chamber 16
2.6. Gene expression analysis 17
2.7. Statistical analysis 17
RESULTS 19
3.1. Measurement of oxygen concentration within the dynamic microfluidic chamber 19
3.2. Cancer cell spheroid formation 21
3.3. NK cell migration assay 23
3.4. Gene expression of functional and chemokine markers in HepG2 cells under normoxic and hypoxic condition 25
3.5. Gene expression of functional and chemokine markers in KHYG-1 cells under normoxic and hypoxic condition 27
DISCUSSION 29
REFERENCES 32
Table 2.1. Primer list of real-time PCR. 18
Figure. 2.1. a Top view of the microfluidic chip. b Bottom view of the microfluidic chip. In the center, cancer cells are inoculated with collagen. On the left, the oxygen-scavenging line is completely separated... 13
Figure. 3.1. a Changes in fluorescence intensity over time measured with a fluorescent microscope inside the chip. b Hourly oxygen concentration over 150 minutes as a function of distance from the central... 20
Figure. 3.2. Before collagen seeding, the HepG2 cells were stained with CellTracker Green. Following seeding, collagen was polymerized for over 2 hours, and the medium was changed daily for 5 days. a... 22
Figure. 3.3. NK cells were labeled with CellTracker Green before flowing through the medium channel. Serum-free RPMI 1640 medium was perfused at a rate of 400 µL/hour using. Under hypoxic conditions,... 24
Figure. 3.4. HepG2 cells within the chip were cultured under hypoxic conditions for 24 hours, followed by retrieval after dissociation from the collagen. The gene expression levels of carbonic anhydrase IX (CA IX),... 26
Figure. 3.5. To analyze changes in gene expression in KHYG-1 cells under dynamic conditions, cells were perfused at a rate of 400 μL/hour and subsequently retrieved. Gene expression levels of functional markers... 28