Cytotolytic activity and characterization of cell-based Biosensor for human natural killer cells against leukemic cells
Date Issued
2010
Date
2010
Author(s)
Chung, Yu-Wei
Abstract
Leukemia, commonly known as blood cancer, is a kind of serious diseases in which cells increase abnormally and invade adjacent tissues. High mortality rate of leukemia indicates its need of effective treatments to cure leukemia[1]. One of promising treatments is cell therapy that requires cell transplantation from healthy donors to patients. The natural killer (NK) cell therapy is chosen due mainly to its natural ability to kill tumor cells with no need of HLA match prior to transplantation. However, natural killer (NK) cells from healthy donors exhibit various levels of cytotoxicity to tumor cells. Effective investigation of the cytotoxicity of natural killer cells to tumor cells remains a great challenge.
This work has developed a cell-based microfluidic device which is capable of investigating the cytotoxicity of natural killer cells against leukemic cells. The whole process of cytolytic activity was observed in real-time under the microscopic system. The lysed k562 cells (leukemic cells) were detected by PhiPhiLux (OncoImmunin, Inc.). All the E/T ratios counted in this work were in a conjugated form, which is the most specific way to count E/T ratios. It was found that the higher conjugated E/T ratio had better cytotoxicity after one hour. Morever, apparent difference in cytotoxocity of the microchannel in gap lengths of 100 μm and 400 μm was found. In a microchannel with a gap length of 100 μm, the cytotoxicity at the conjugated E/T ratio of 1,2,3,4 were 0.75, 0.8, 1, 1, respectively. In a device with a gap length of 400 μm, the cytotoxicities were 0.63, 0.33, 0.75, 0.67, respectively. The mean value of cytotoxicity over all conjugated E/T ratio was higher than 0.6. The total conjugated E/T ratios formed in three microchannels were between 2.1 and 2.3. The averaged cytotoxicities were 0.85, 0.72 and 0.63 in 100 μm, 200 μm and 400 μm channel, respectively. By comparing those three microfluidic channels, it was found that the small size channel formed the higher density of the cells and thus performed the higher cytotoxicity.
Unlike the conventional cell culture experiments in dishes with large amounts of cells, this work provides a device that needs very few cells for manipulation and significantly reduces the consumption of culture medium. The microscopic system also provides an in-depth understanding of cell interaction and cytotolytic activity. This microfluidic device exhibits the capability and potential for effective investigation of natural killer cells (NK cells) against tumor cells (K562 cells).
Subjects
leukemia
natural killer cell
leukemic cell
cell-based biosensor
cytotoxicity
conjugated E/T ratio
SDGs
Type
thesis
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