Abstract
BackgroundThe use of acoustic forces to manipulate particles or cells at the microfluidic scale (i.e. acoustophoresis), enables non-contact, label-free separation based on intrinsic cell properties such as size, density and compressibility. Acoustophoresis holds great promise as a cell separation technique in several research and clinical areas. However, it has been suggested that the force acting upon cells undergoing acoustophoresis may impact cell viability, proliferation or cell function via subtle phenotypic changes. If this were the case, it would suggest that the acoustophoresis method would be a less useful tool for many cell analysis applications as well as for cell therapy.MethodsWe investigate, for the first time, several key aspects of cellular changes following acoustophoretic processing. We used two settings of ultrasonic actuation, one that is used for cell sorting (10 Vpp operating voltage) and one that is close to the maximum of what the system can generate (20 Vpp). We used microglial cells and assessed cell viability and proliferation, as well as the inflammatory response that is indicative of more subtle changes in cellular phenotype. Furthermore, we adapted a similar methodology to monitor the response of human prostate cancer cells to acoustophoretic processing. Lastly, we analyzed the respiratory properties of human leukocytes and thrombocytes to explore if acoustophoretic processing has adverse effects.ResultsBV2 microglia were unaltered after acoustophoretic processing as measured by apoptosis and cell turnover assays as well as inflammatory cytokine response up to 48 h following acoustophoresis. Similarly, we found that acoustophoretic processing neither affected the cell viability of prostate cancer cells nor altered their prostate-specific antigen secretion following androgen receptor activation. Finally, human thrombocytes and leukocytes displayed unaltered mitochondrial respiratory function and integrity after acoustophoretic processing.ConclusionWe conclude that microchannel acoustophoresis can be used for effective continuous flow-based cell separation without affecting cell viability, proliferation, mitochondrial respiration or inflammatory status.
Highlights
The use of acoustic forces to handle particles and cells in microfluidic systems is gaining increased attention [1]
In view of its high reproducibility, reliability and the fact that this technology can be applied to most cell types, acoustophoresis holds great promise as a cell manipulation technique in several research and clinical settings [13]
Microparticles were used instead of cells do to their homogenous size distribution, which allows for a better measurement
Summary
The use of acoustic forces to handle particles and cells in microfluidic systems (i.e. microchannel acoustophoresis) is gaining increased attention [1]. The application in which the acoustophoresis method can be used include particle manipulation [2,3], depletion [4], washing [5,6,7], fractionation [8], rare event sorting [9,10], concentration [11] and cell cycle synchronization [12] This novel cell manipulation technique is label-free and enables separation by unique cell properties, e.g. compressibility. Hultstrom and colleagues [17] as well as Evander et al [18], using cos-7 cells from fetal monkey kidney and rat neural stem cells respectively, studied cell viability in an acoustic trap. If this were the case, it would suggest that the acoustophoresis method would be a less useful tool for many cell analysis applications as well as for cell therapy
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