Cell Unclumping Assay in Drug Discovery: Sodium Sulfate

Abstract

Clumped cells are generally more dangerous than single cells in cancer spread, thrombocytosis and biofilm infectivity. Here a simple 3 component direct kinetic assay is used to examine 2 reagents for anti‐clumping activity using a Prefer (Anatech Ltd., Battle Creek, MI) fixed yeast (Saccharomyces cerevisiae) model that has been recently described by us in detail using other reagents (Amer J Applied Scientific Research 5:28–34 March 2019). In 1212 trials by 17 investigators sodium sulfate and sodium bicarbonate (1–3 mg per ml deionized water) were examined by measuring percentage single cells, number of clumps and number of cells per clump over a 60 min time course, with standard deviations and t‐tests to determine any significant differences between controls and experimentals. Both reagents showed sometimes inconsistent unclumping activity especially in magnitude of effects. When percentage of single cells increased, clump number and/or number of cells per clump generally decreased, helping to validate the assay. An example of these findings in 60 trials at 60 min with 1–3 mg sodium sulfate per ml deionized water: 1 mg 15% increased singles (p<0.01), 29% decreased clumps (p<0.01), 11% decreased cells per clump (p>0.05); 2 mg 12% increased singles (p<0.01), 20% decreased clumps (p<0.01), 30% decreased cells per clump (p<0.01); 3 mg 27% increased singles (p<0.01), 36% decreased clumps (p<0.01), 28 % decreased cells per clump (p<0.02).Here sodium sulfate showed promise as an anti‐cell‐clumping reagent together with sodium citrate reported previously (Amer J Applied Scientific Research 5: 28–34 March 2019). Sodium citrate is a known human anticoagulant (Platelets 29 (2018) 21–26) independently identified with this assay, helping to validate the assay for drug discovery applications.Support or Funding InformationSupported by California State University, Northridge, Department of Biology and Center for Cancer and Developmental Biology