Reprogramming of fibroblast cells to totipotent state by DNA demethylation

Abstract

Many attempts have been made to induce high-quality embryonic stem cells such as pluripotent stem cells and totipotent stem cells, but challenges remain to be overcome such as appropriate methods and sources. Demethylation of the genome after fertilization is an important step to initiate zygote gene activation, which can lead to the development of new embryos. Here, we tried to induce totipotent stem cells by mimicking DNA demethylation patterns of the embryo. Our data showed, after induction of DNA demethylation via chemicals or knockdown of Dnmts, cells positive for Nanog, and Cdx2 emerged. These cells could differentiate into the pluripotent and trophoblast lineage cells in-vitro. After transferring these cells to the uterus, they can implant and form embryo-like structures. Our study showed the importance of DNA demethylation roles in totipotent stem cell induction and a new and easy way to induce this cell type.

Epigenetic reprogramming in the transition from pluripotency to totipotency

“…Recent studies show that the regulation of DNMTs and TET may ultimately determine the dynamics of DNA demethylation in totipotent states (Eckersley-Maslin et al, 2018;Ghazimoradi et al, 2023) (Figure 3c). Mapping genomic DNA hypomethylation patterns reveals hidden rules of epigenetic reprogramming during the transition from pluripotency to totipotency.…”

Enhancing Cell Growth with PAN/PVA-Gelatin 3D Scaffold: A Novel Approach using In-situ UV Radiation Electrospinning and Plasma Treatment

“…The mFib-2 cells, which are classi ed as "Normal" broblast-like cells, were isolated using the methodology outlined in prior research 27 . Then they were cultured in a medium containing 78.5% DMEM high glucose, 10% FBS, and 1.5% antibiotics (penicillin and streptomycin) at 37°C and 5% CO 2 .…”