Development of a Microdilution Device with One-step Dilution of Cytochalasin-B for Treating ORL-48 Cancer Microtissues

摘要

Mixing and dilution are essential procedures in pharmaceutical operation to aliquot two or more components serially to produce less concentrated and well mixed solutions. However, conventional serial dilution method used in laboratory is tedious and utilized large quantity of plasticwares. In this study, a two-tier microdilution device with two inlets and four outlets was designed, simulated, and prototyped to dilute Cytochalasin-B (CB). Using the microdilution device, CB in linear concentration gradients were produced based on one-step dilution method. The different concentrations of CB were applied to treat monolayer and microtissues of ORL-48 cells. The morphological responses, cell viability and cell proliferation of ORL-48 monolayer cells (2D) and microtissues (3D) treated in four different CB concentrations were investigated via phase contrast microscopy, live/dead stainings, and Alamar Blue (R) assay, respectively. The results showed that 2D ORL-48 cells were morphologically affected but 3D ORL- 48 cells stayed viable and proliferative after treated with similar concentrations of CB. The microdilution device enables serial dilutions to produce reagents in linear concentration gradients via a one-step dilution method.