Cell-based Microfluidic Device for Cytotoxicity and Real-time Investigation of Primary Human Natural Killer Cells against Leukemic Cells
Date Issued
2012
Date
2012
Author(s)
Lin, Yi-Wen
Abstract
Natural killer cell transplantation therapy is potential to cancer treatment. It is able to killer cancer cells directly and has no need of haploidentical matched donor before transplantation. Since natural killer cells from different donors perform different cytotoxicity against cancer cells, it is significant to choose an appropriate donor who is suitable for each patient. However, the conventional method for determination of cytotoxicity took large amounts of cells and could not provide image evidence of cell-cell interaction process. As a result, we developed a new appropriate assessment to select multiple healthy donors.
In this study, we designed a cell-based microfluidic device which was capable to integrate a real-time observation system. The effector cells we used were primary natural killer cells isolated from peripheral blood. The target cells were leukemic cell line K562. The cytotoxic assay performed by micro device needed only small amount of samples. And the results showed that the micro device is able to replace conventional cytometry to determine the cytotoxicity of natural killer cell. Furthermore, this study adopted primary natural killer cells from three healthy donors. Therefore, the experimental results provide the basis for a suitable donor in transplantation.
This study developed a micro device applied on research of suspension cells. The micro device was fabricated by MEMS technique and easily integrated with real-time observation system. With microfluidic manipulation and the designed gap structure, the micro device could trap dozens of cells. The liquid pressure difference between central and side channels were controlled to maintain cell docked in the gap and complete cell identification of apoptosis. The cytotoxicity results from the micro device were consistent with that from conventional cytometry. Moreover, the cytotoxicity of NK cells from donor A, B and C which were cultured in vitro for 14 days were 70.07%,66.53% and 61.01%, respectively. After culturing 21 days in vitro, the cytotoxicity of NK cells were not growing significantly. We proved that the cell-based biosensor reduced the consumption of cell sample from the number of 2×105 to 101 for detection and present the capability and potential for application of natural killer cell transplantation.
Subjects
natural killer cell
microfluidic
biosensor
MEMS
real-time observation
SDGs
Type
thesis
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