Comparing Effectiveness of Nanog/Pou5f3 and SOX2/Lin28a in Pluripotent Stem Cell Induction of Leghorn Chicken Fibroblasts

Abstract

Pluripotent stem cells have incredible differentiation potential, making them ideal candidates for various therapeutic and research applications. One particularly interesting application involves the use of stem cells from endangered species for conservation purposes. Naturally harvesting pluripotent stem cells, however, involves significant health risks and is, therefore, avoided in the context of conservation. Induced Pluripotent Stem Cells (iPSCs) can serve as a replacement due to their minimally-invasive potential. Previous literature tends to utilize a large and possibly redundant amount of genes to induce pluripotency. While effective, issues may arise regarding vector capacity and potential transcription factor conflicts. Our study sought to elucidate the necessary genes to induce pluripotency in avian fibroblasts by comparing the effectiveness of two common gene combinations, Nanog/Pou5f3 and Sox2/Lin28a, using the Leghorn Chicken as a model organism. Due to the essential nature of genes Nanog/Pou5f3, we hypothesized that they would be more effective at inducing pluripotency than genes Sox2/Lin28a. Using RT-qPCR, we quantified the gene expression of pluripotent maintenance genes and found that the overexpression of Sox2/Lin28a upregulated Sall4 and Klf4 more effectively than Nanog/Pou5f3. We also found that the presence of Nanog/Pou5f3 may inhibit the upregulation of Sall4 via Sox2/Lin28a. Finally, we present evidence of Klf4 upregulation via both Nanog/Pou5f3 and Sox2/Lin28a, a potentially novel mechanism.CFAES Small Grant AwardA one-year embargo was granted for this item.Academic Major: Animal Science