A Comparative Study Of Cell Culture Conditions During Conversion From Primed To Naive Human Pluripotent Stem Cells

Abstract

Since the successful reprogramming of human somatic cells into induced pluripotent stem cells (hiPSCs), stem cells have opened up a large field of research to generate new therapies due to their ability to differentiate into any cell type. Embryonic stem cells (ESC) are pluripotent cells which give rise to all somatic cell types in the embryo. Their self-renewal ability and plasticity allows for in vitro generation of many distinct cell types, raising new challenges for regenerative medicine and therapies. The “naive” state of cell pluripotency is the result of cells that come from the preimplantation epiblast in vivo. This state was observed in mouse embryonic stem cells and was characterized by a high proliferation and differentiation capacity as well as a global DNA hypomethylation. Human embryonic stem cells (hESCs) are derived from the inner cell mass of preimplantation embryos and correspond to a later stage called “primed” stage of embryonic development. The conversion of this “primed” human embryonic stem cells (hESCs) to a “naive” state is desirable as their characteristics would facilitate many techniques such as gene editing and a more efficient differentiation. The main objective of this conversion is to facilitate the application of cell therapies to be able to use them in clinical treatments to model different diseases, such as human primary immunodeficiencies related to NK cell defects. In the present study, the main objective is to compare and evaluate different culture conditions for conversion from primed to naive state of an hES cell line called ES-2. The different culture conditions are based on different conversion media (Gafni, Fine and T2iLGö) in both feeder and feeder-free cells conditions.