Cell passage process in a microchip
From LabAutopedia
Cell passage process in a microchip
Elzbieta Jedrych, Michal Chudy, Artur Dybko, Zbigniew Brzozka
Warsaw University of Technology, Department of Microbioanalytics,
Noakowskiego 3, 00-664 Warsaw, Poland
ejedrych@ch.pw.edu.pl
A microsystem for cell culture and analysis should provide growth and proliferation of cells, control of cells migration, adhesion and optimal oxygenation and flow of substances (i.e. medium, trypsin). Passage process is important in cell culture and is critical for maintaining undifferentiated and stable cell lines. Passage assures proper conditions for growth and proliferation of cells. Without that, death of cells can be caused by nutrient amount decrease in the culture medium, and increase of toxic metabolism products, so the passage process is essential for continuous cell culture. Passage process in a microsystem is a new achievement of microtechnology.
The aim of the studies was to design a miniaturized system dedicated for cell culture, that enables a passage process. The hybrid microdevice glass/PDMS consisted of a microchannels network for medium dosing and the microchambers for cells’ seeding and culture. The A549 human lung carcinoma cells were docked in the microchambers and cultured in the microdevice for 5 days. The growth and proliferation was possible due to the device sterility. The density of cells seeded in microdevice and medium dosage was also elaborated. After 5 days, when cells reached the confluence state in the microchambers, cells’ passage was performed according to the procedure applied for standard scale cell culture. The passage process was based on partial cells’ flushing with trypsin EDTA-solution. The trypsinization step caused partial cells’ detaching and after that of 60-70% cells were removed from the device. After passage good cells’ adhesion and the ability to further growth was observed. Regular passage process was essential for maintaining a healthy cell line.
The successful attempts of cell passage in the designed microdevice were performed while cells confluence was achieved. The novel method of cells’ passaging in our microsystem was highly effective and repeatable and the passage process gave possible continuous culture in microfluidic device, so this step decreases costs and waste amount, which is important for the natural environment protection. The presented microsystem can be applied in high throughput cell based assays. Moreover, it is possible to test in microscale, how passage numbers influence on alterations in cell morphology, response to stimuli, growth rates, protein expression and cell analysis.
