cellometer vision cell counter  (Revvity)

Journal: Molecular Therapy Oncolytics

Article Title: Acquired chemoresistance can lead to increased resistance of pancreatic cancer cells to oncolytic vesicular stomatitis virus

doi: 10.1016/j.omto.2021.11.019

Figure Lengend Snippet: Viral replication kinetics (A) C and GR cell lines were either mock treated or infected with VSV-ΔM51 at MOIs of 1.0, 0.1, or 0.01. GFP fluorescence was measured over time from 1 to 72 h p.i. Control and GR cell lines 1–3 are combined for each MOI. (B) Cell viability of C and GR cells 70 h after infection at different MOIs. Control and GR cell lines were either mock treated or infected with VSV-ΔM51 at MOIs of 1.0, 0.1, or 0.01. Cell viability was measure 70 h p.i. using a WST-8 cell viability assay. The figure represents data from three independent experiments. (C and D) Percent of infected C and GR cells to determine differences in ability of VSV to spread. Control and GR cells were infected with VSV-ΔM51 at MOIs of 1.0, 0.1, and 0.01. Samples were collected at 13 and 24 h p.i. Samples were trypsinized and the percent of GFP-positive cells was determined by using a Nexcelom Cellometer Vision fluorescent cell counting system. (E and F) Virus particle production: one step and multi-step virus kinetics in C and GR cells. Control and GR cells were infected with VSV-ΔM51 at an MOI of 0.01 (E) or MOI 10 (F). Wells were washed after initial 1-h infection and fresh media was added to all wells. Supernatant was collected at 1, 12, 24, 48, 72, and 96 h p.i. Virus titers were determined for each time point using standard plaque assay on BHK-21 cells. The data points and error bars shown represent the means and SEM of the means, respectively (some error bars are too small to be seen in the figures). Results were analyzed to determine significance using a Student's t test or one-way analysis of variance with a Sidak's multiple comparison test at a 95% confidence interval for comparison between each condition. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. Control and GR cell lines 1–3 are combined for each MOI.

Article Snippet: Samples were trypsinized and the percent of GFP-positive cells was determined by using a Nexcelom Cellometer Vision fluorescent cell counting system. (E and F) Virus particle production: one step and multi-step virus kinetics in C and GR cells.

Techniques: Infection, Fluorescence, Control, Viability Assay, Cell Counting, Virus, Plaque Assay, Comparison