Vi cell cell counter for cell viability analyzer

In order to study the impact of miRNAs on cell proliferation, hASCs were transfected with miR mimic, miR inhibitor, or negative control. Cell proliferation was assessed for each miRNA in three independent biological replicates. For all experiments, transfected cells were seeded at a fixed density of 2 Â 10 4 cells per well right after transfection (day 0) in primary hASC culture medium consisting of DMEM-low glucose/HAM's F-12 (GE Healthcare, Vienna, Austria) medium supplemented with 10% fetal calf serum (FCS; Sigma-Aldrich, Munich, Germany), 4 mM Lglutamine, and 1 ng/mL recombinant human basic fibroblast growth factor (rhFGF; R&D Systems, Minneapolis, MN, USA). Seventy-two hours later (day 3), the number of viable cells/mL was automatically assessed using the Vi-CELL Cell Counter for Cell Viability Analyzer (Beckman Coulter, Brae, CA, USA), which operates based on the Trypan Blue Dye exclusion method. In order to account for background proliferation changes induced by the transfection process itself and to allow for interdonor comparability, we calculated the fold change of viable cell density on day 3 in miR/miR inhibitor-transfected cells compared to negative control-transfected cells.