Evaluating the Effects of Cell Sorting on Gene Expression
- J Biomol Tech. 2020 Jun 22:jbt.20-3103-004. doi: 10.7171/jbt.2020-3103-004.
- 1. Stowers Institute for Medical Research, Kansas City, Missouri, USA.
- 2. Cincinnati Children's Hospital, Cincinnati, Ohio, USA.
- 3. University of Vermont Cancer Center, Burlington, Vermont, USA.
- 4. Center for Functional Genomics, University at Albany, Albany, New York, USA.
- 5. Dartmouth College, Hanover, New Hampshire, USA.
- 6. University of Rochester Medical Center, West Henrietta, New York, USA.
- 7. New York University Langone Medical Center, New York City, New York, USA.
- 8. University of Pittsburgh Cancer Institute, Pittsburgh, Pennsylvania, USA.
- 9. The Scripps Research Institute, San Diego, California, USA.
- 10. Department of Pediatrics and Cincinnati Children's Hospital, University of Cincinnati Medical Center, Cincinnati, Ohio, USA; and.
- 11. West Virginia University, Morgantown, West Virginia, USA.
Cell sorting is a commonly used technology to isolate highly purified cell populations for downstream applications. Because the sorted cells are destined for further analysis, i.e., gene expression assays or functional assays, ensuring that the sorting process itself has little effect on the cells is of utmost importance. Previous studies examining the effects of sorting on cellular function have primarily focused on a specific cell type or condition. One of the goals of the Flow Cytometry Research Group of the Association of Biomolecular Resource Facilities is to establish best practice guidelines for cell sorting conditions that minimize cell stress, perturbation, or injury to the sorted cell population. In this study, the effects of nozzle size, sample pressure, UV exposure, and instrument type were evaluated for their effects on gene expression and cell cycle using both established cell lines and primary cells across several flow cytometry shared facilities. Results indicate that nozzle size and pressure, as well as UV exposure and instrument type, have only minor effects on gene expression, which were diminished by subsequent culturing of the sorted cells. In this assessment, these data demonstrate that cell sorting itself, regardless of instrumentation used, has minimal effects on downstream cellular applications.