A Real-Time Investigation on Cytotoxic Assay of Cytokine-activated Human Natural Killer Cells against Cancer Cells in a Microfluidic Device by Inverted Point Laser Scanning Confocal Microscopy
Date Issued
2014
Date
2014
Author(s)
Lu, Hsiang-Jen
Abstract
Leukemia, known as blood cancer, occurs due to DNA variation of human bone marrow stem cells, making too many immature white blood cells. A large number of these immature white blood cells present in the blood seriously affect the normal function of blood cells. Due to little side effect of natural killer cell, natural killer cell therapy has been chosen as an auxiliary strategy in post chemical treatment for good quality of life.
In this study, a microfluidic cell-trapped device was designed and manufactured by MEMS technique. With the design of narrow gaps between channels, suspension cells were trapped by a fluidic pressure drop between channels to be capable of trapped cell monitoring or observation under a microscopy. As a result, cell-cell interaction and cell apoptosis processes can be investigated with this device under a currently used fluorescence microscopy. In this study, cytokine interleukin 12 (IL 12) has been chosen to activate human natural killer cells in killing cancer cell (K562) process. With various ratios of effector cells to target cells (E/T ratio) in a cell-trapped device, a microfluidic cytotoxic assay was analyzed under a fluorescence microscopy. The cytotoxic results of the present device showed an excellent agreement with those by the conventional flow cytometry. The merit of the present device shows a feasible cytotoxic analysis using few cells in an order of 102 instead of around 106 cells for analysis in conventional flow cytometry. Additionally, the cytotoxicity of NK cells against cancer cells (K562) was found in a range from 49.7% to 79.2% over time periods in a cytokine IL-12 cell activation. Moreover, trapped in a device with an inverted high-resolution confocal microscopy by scanning fluorescent staining cell samples, the on-chip NK and cancer mixed cells in a cytokine-activated medium fluidics were kept monitoring in a long-term tracking of cell killing activities. Cell morphology was found to gradually change over time. Cell apoptosis process of cell shrinkage, membrane budding, and apoptosis bodies were firstly found with the present device in this experiment.
Lastly, the on-chip determination of cell killing process and activity has been firstly achieved by this present device.
Subjects
人類自然殺手細胞
微流體
生物晶片
微機電製程
共軛焦顯微鏡
SDGs
Type
thesis
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