Anna Marie Jones, Cameron Rankin
Faculty Sponsor: David Wessner
A persistent problem faced when trying to determine cell viability is determining between the plethora of options offered to essentially arise to the same conclusion. We evaluated the effectiveness of various types of cytotoxicity assays used commonly in mammalian cell culture. Cytotoxicity assays are a commonly used in component of the undergraduate microbiology teaching laboratories to in cell cultures to examine cell viability after exposure to cytotoxic compounds. We aim to identify the most efficient effective assay for an undergraduate teaching laboratory course through using four common in vitro cytotoxicity assay types, evaluating them on the criteria of accuracy in measuring cell death, cost of assay, and feasibility of use in an undergraduate setting. For each assay, 96 well plates, were seeded with either Madin-Darby canine kidney (MDCK) cells or murine fibroblast (L929) cells, and then left overnight to incubate. The next day, cells were then exposed over for twenty-four hours to two-fold dilutions of 95% ethanol to induce cell death. Cell viability was then assessed using neutral red assays, crystal violet assays, lactate dehydrogenase (LDH) assays, and trypan blue assays. We found that a trypan blue assay utilizing acetic acid consistently outperformed the other assays in accuracy of recording cell viability. Also, Using ttrypan blue offered a relatively easy, affordable, and time efficient assays, leading to our recommendation of a trypan blue viability stain as an ideal methodology to be used in an undergraduate course. With all of these criteria, we suggest trypan blue viability assays are the ideal assays to teach in undergraduate courses.