DNA methylation abnormal accumulation during cloned TSCs maintaining [RRBS]

Trophoblast stem cells (TSCs) are derived from the trophoectoderm of blastocysts and maintain ability to self-renewal and differentiation. TSC is a good model to research placenta development in vitro. It will contribute to understanding and improving cloned placentomegaly to compare the transcription and methylation between cloned and natural fertilized embryos throughout TSC derivation process. In the present study, we used SCNT (NT) and SCNT with HDACi treatment (SNT) as cloned groups and natural fertilized (NF) embryos to derive TSCs and chosed 5-time points to peform RNA-seq and RRBS. We found only NT got a barriar in TSC maintenace and both cloned groups exhited abnormal accumulating DNA methylation and it might be resiponsible for some malformations of cloned placentas. Overall design: We generated embryos from 3 different sources natural fertilization (NF), somatic nuclear transfer (NT), and somatic nuclear transfer with HDACi Scriptaid treatment (SNT) and chosed 5-time points during the TSC maintaining, E3.5 blastocyst trophectoderm (TE3.5), E4.5 blastocyst trophectoderm (TE4.5), outgrowths at half-time progression (outgrowth), passage 1 TSCs (TS_P1), and passage 3-4 TSCs (TS_Pn) to perform RNA-seq and RRBS. For each sample, we performed several independent replicates.