A Real-Time Investigation on Cytotoxic Assay of Primary Human Natural Killer Cells against Leukemic Cells in Microfluidic Device
Date Issued
2015
Date
2015
Author(s)
Wang, Jian-Jhih
Abstract
In recent years, natural killer cells (NK cells) therapy was used to treat leukemia in increasing clinic cases, due to its less side effects that patients can maintain good quality of life. Since NK cells from different individuals exhibit different performances of cytotoxicity, it is important to choose great cytotoxicity from different donors. In tradition, cytotoxicity detection is performed by using flow cytometry; however, it requires a large numbers of cells (5×105~106) which show a challenge in acquiring primary NK cells of patients in clinic. In this study, we used MEMS technique to fabricate microfluidic device which allows suspension cells to be trapped by the method of microchannel design and pressure difference manipulation of liquid. By integrating the microfluidic device with fluorescent staining techniques and inverted fluorescent microscope, the real-time observation system was established for the detection of cytotoxicity. By using the microfluidic device we designed to detect the cytotoxicity of NK cells against leukemia, the result showed that the cytotoxicity is higher when effector to target cell ratio (E/T ratio) is greater. For the present microfluidic device to be used practically in clinic, we obtained primary NK cells from donors, and detect the cytotoxicity once a week. After ex-vivo culture for fourteen days, the cytotoxicity saturated, which perform 63.29 %, 73.90 %, 56.38 %, 75.68 %, respectively. Furthermore, we also compared the cell killing performance of the microfluidic device with flow cytometry, and the results of both methods were comparable. This study successfully analyzed the cytotoxicity of a few NK cells (101~102) by using microfluidic device. Furthermore, the result of the microfluidic device was consistent with the flow cytometry which commonly require a great number of cells to perform detection.
Subjects
primary natural killer cells
microfluidic device
MEMS
real-time observation system
Type
thesis
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