Investigations of Methods for Non-Viral Generation of Bovine Cells to Pluripotency, With a View on Potential Use in Reproductive Technologies

Abstract

Spermatogonial stem cells (SSCs) from livestock species have the potential to be used for reproductive technologies and for the production of transgenic animals. The isolation of a relatively pure population of SSCs from livestock species has proven difficult; however, the recent advances in the production of induced pluripotent stem (iPS) cells may provide an alternative source of SSCs through the differentiation of iPS cells toward the germline. The general aim of this thesis was to produce bovine iPS cells that may be differentiated toward the germ line for potential use in reproductive technologies such as germ cell transplantation. Due to difficulties in isolating a pure population of bovine SSCs, methods to improve the enrichment of these cells are of interest in order to improve the success of bovine germ cell transplantation. A number of different methods to enrich bovine spermatogonia were trialled to determine which of the method for enrichment was most effective. The combination of enrichment by differential plating followed by separation of cells by discontinuous Percoll gradient centrifugation, was found to isolate the most enriched population of un-differentiated spermatogonia.