In-house polydimethylsiloxane microfluidic device on single cell trapping and culturing of Leukemia cell line: comparison between conventional and conventional with centrifugal methods

Abstract

To develop a suitable model to study biological and characterization of Cancer stem cells (CSCs), single cell trapping and culturing using microfluidic techniques have become a high-through put system that can manage and control microenvironment and practical intracellular analysis. The aim of this study was to develop and validate a microfluidic device for single cell trapping and culturing using the leukemia cell line as a cell model. Comparison between conventional and conventional with centrifugal methods was also analyzed. To compare trapping efficacy between these 2 methods, leukemia cell concentration of 1.5×106 cells/ml was loaded with a flow rate of 0.25 ml/h. In the conventional with centrifugal method, the chip was centrifuged twice with 1,000 rpm/rcf for 2 mins. From the results, the conventional method with centrifugal method showed significantly higher numbers of trapping microwells than the conventional method in all trapping manners. To examine culture efficacy, the cells were cultured in microfluidic devices and stained with trypan blue dye at 24, 48 and 72 h for cell viability study. The results showed that the conventional with centrifugal method showed a higher percentage than the conventional method at 48 h and significantly at 72 h. Both methods showed a high percentage of viable cells at 24 h. However, percentages at 48 and 72 h were significantly decreased. In summary, our developed microfluidic device was successful in trapping single cell pattern and short-term culture of single cell of the leukemia cell line. In addition, administration of an additional centrifugal method could be used to increase cell trapping efficacy when compared to the conventional method.