| Objectives: In vitro experiment plays an important role in biomedical research.Studies on cell motility, cell-cell interactions and other cell life activities wouldcontribute to revealing further exploration of disease pathogenesis and the intrinsic lawof biological phenomena. Such in-vitro studies require much closer simulation of invivo environment to achieve more authentic outcomes. However, the traditional singlecell culture can’t meet the above requirement. Thus, there has been an urgent need formultiple cells co-culture in the field of biomedical science. At present, one keyproblemthe co-culture technology facing is the subsequent cell isolating for thedetection in follow-up process. Therefore, cell sorting has increasingly become themost important task for co-culture. Many studies have indicated the diameters ofdifferent cells vary a lot which can be the classification rules for cell sorting. This studyintended to construct a microfluidic platform driven by an electric field for cell sorting.In that case, the human lung adenocarcinoma cells A549and human bronchialepithelial cells16HBE can be co-cultured on this platform. Then relevant study can becarried out to detect the epithelial-mesenchymal transition (EMT) on these two celllines.Methods: A microfluidic device driven by electric was designed and constructedfor cell sorting. The device was consisted of a microfluidic chip, an electronicdifferential amplifier, a LabView system for data processing and analysis and otherassays. The mixture suspension of A549cells and16HBE cells was prepared and theninjected into the channel of the chip. Due to the difference sizes of these two cells, theyproduced different signals when entering the testing region. Program was set on thebasis of signal difference to capture cells and control the voltage conversion of Double Pole Double Throw (DPDT) so that it can produce instant low voltage direct currentpower for the isolating of target cells. After being sorted, the vitality of the isolatedcells was determined via Trypan blue staining. Then these target cells were put on themicrofluidic platform for continuous cultivation. The apoptotic rate within certainextend was determined via Hoechst33342/PI double staining. Transforming growthfactor beta (TGF-β1) was added into the chip for inducing an EMT event in both A549and16HBE cell lines. The differences of EMT between single culture and co-culture ofthese two cell lines were detected and analyzed.Results: We successfully developed a microfluidic platform driven by an electricfield for cell sorting. Target cells sorting based on sizes was implemented. The largerA549cells were enrolled into the collecting channel in the upper part of the platformwhile the smaller16HBE cells into the lower part. The cell vitality presented wellafter cell sorting according to Trypan blue staining, without obvious apoptosis. Theapoptotic index of examined cells was lower than10%after a48-hour continuouscultivation.When cultured separately, the time for TGF-β1-induced EMT for humanbronchial epithelial16HBE cells and human lung adenocarcinoma A549cells was36hours and24hours, respectively, indicating there was a12hours prolonging for16HBEcompared that to A549cells. But when these two cells were co-cultured, no matter for16HBE or A549cells, the time for EMT was significantly shorter than that of singleculture mode.Conclusion: The present study successfully designed and constructed amicrofluidic platform that can be used for cell culture and detection cell vitality, cellapoptosis and the inducible EMT. Such a technology can provide a novel platform forcell sorting and biological research on EMT. |
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