Implications of culture media changes in embryonic stem cell properties

• Autor(a) principal: Mateus, Francisca Cazaux
• Data de Publicação: 2022
• Tipo de documento: Dissertação
• Idioma: eng
• Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
• Texto Completo: http://hdl.handle.net/10773/35648
• topic: Embryonic stem cells; Cancer stem cells; Pluripotency; DNA methylation

Embryonic stem cells are pluripotent stem cells derived from the inner cell mass of preimplantation embryos. These cells can be maintained in vitro under carefully maintained culture conditions. The discovery of these cells lead to great advances not only in the stem cell biology field but in so many others since they can be differentiated into different cell types. In this project, we evaluated how different culture conditions affect distinct properties of the TNG-A cell line, which is a transgenic mouse embryonic cell line with a green fluorescent protein (GFP) insert at one of the Nanog alleles. We specifically assessed how the optimal (DMEM LIF/2i) and sub-optimal (DMEM LIF) culture conditions affect the pluripotency status of these cells, the DNA methylation marks, and lastly, the similarities of cancer stem cells and TNG-A in terms of protein expression. We saw that the two inhibitors (2i) are important for the maintenance of pluripotency and confirmed that they are potent methylation inhibitors leaving the cells hypomethylated. Moreover, we confirmed that TNG-A are more alike cancer stem cells when they are cultured without the two inhibitors and that two proteins involved in the metabolism, Psph and Glod4, show a similar expression in these two types of cells. These results showed the importance these two inhibitors have for the maintenance of embryonic stem cells properties and that TNG-A show high plasticity and sensitivity to external signals.