Abstract

ABSTRACT OF THE THESISSingle-Cell Neurosphere Assay: A Method to Quantify Neural Stem Cells Clonogenicity on a Single Cell LevelbyGivon John SanatiMaster of Science in BiologyUniversity of California, San Diego, 2016Professor Alexander Groisman, Chair Professor James Kadonaga, Co-ChairThe progression of stem cell therapy continues to be interrupted by the absence of consistent stem cell clonogenicity protocols. Previously administered clonogenicity assays demonstrate some promise; however, they are restricted to specific cell types. We take advantage of a microfabricated approach to study neural stem cell clonogenicity on a single cell level. We engineer a microarray platform using silicone matrix technology to study clonogenicity. In addition, we use image cytometry to quantify functional characterization of clonogenicity. Lastly, we explore and compare known stem cell biomarkers to further validate our functional assay. We found that early passaged neural stem cells demonstrate higher neurosphere formation capacity as compared with later passaged neural stem cells. To further confirm this outcome, we identified CD133 as a known clonogenic biomarker for neural stem cells. We determined a possible correlational relationship between CD133 and neural stem cell clonogenicity. We establish and confirmed a novel approach to study clonogenicity. This approach will allow for a more reliable method to compare and study stem cell potency.