Effect of different culture media on epigenetic marks in bovine fibroblasts

Abstract

The epigenome reprogramming mediated by somatic cell nuclear transfer (SCNT) created a huge expectation during the nineties, with several applications such as production of transgenic animals and cloning of high-genetic merit animals. The birth of Dolly was possible due to the perfect cellular synchronization, who was achieved through serum deprivation during the cell culture, inducing the cells to the quiescent state. Around twenty years has passed since Dolly's birth, and the wide use of SCNT has been impeded due to the poor outcome. These is believed to be caused by faulty epigenetic reprogramming of the donor cells. Among the factors responsible by the epigenome alterations commonly observed in in vitro produced embryos, the fetal calf serum is judged to be the main player. Nutritional factors can induce modifications on epigenetic marks, consequently affecting the nuclear reprogramming process. Based on this, the hypothesis of this proposal is that the epigenetic marks from somatic cells utilized as nuclear donors (fibroblasts) on SCNT procedure changes in response to the cell culture environment or stage of cell cycle. For this reason, here we propose to investigate the effects of several kinds of culture media on epigenetic modifications in bovine fibroblasts. We expect with this project contribute to the basic sciences, aiming understand does the organismal somatic cell epigenome can be modulated in response to variations in the cell culture environment.