Project description:Current human pluripotent stem cells lack the transcription factor circuitry that governs the ground state of mouse embryonic stem cells (ESC). Here we report that short-term expression of two components, NANOG and KLF2, is sufficient to ignite other elements of the network and reset the human pluripotent state. Inhibition of ERK and protein kinase C signalling sustains a transgene-independent rewired state. Reset cells self-renew continuously without ERK signalling, are phenotypically stable and karyotypically intact. They differentiate in vitro and form teratomas in vivo. Metabolism is reprogrammed in reset cells with activation of mitochondrial respiration as in ESC. DNA methylation is dramatically reduced and transcriptome state is globally realigned across multiple cell lines. Depletion of ground state transcription factors, TFCP2L1 or KLF4 has marginal impact on conventional human pluripotent stem cells, but collapses the reset state. These findings demonstrate feasibility of installing and propagating functional control circuitry for ground state pluripotency in human cells. DNA methylation analysis in Conventional and Reset human embryonic stem cells by whole genome bisulfite sequencing, in triplicate, using the Illumina platform
Project description:Transcriptomic analysis of mouse embryonic stem cells (mESC) maintained in the pluripotent state or allowed to differentiate for 3 or 7 days by leukaemia inhibitory factor (LIF) withdrawal.
Project description:Embryonic stem cells are pluripotent and possess the ability to differentiate into numerous lineages during the developmental process. In similarity to embryonic stem cells, human induced pluripotent stem cells (iPSCs) possess the potential to differentiate into multiple lineages making them an excellent research tool. We generated iPSCs from multiple donors and also differentiated iPSCs from these donors into human neural progenitor cells (NPCs). We used human transcriptome arrays to detail the programme of gene expression underlying NPC induction and identified distinct classes of up-regulated genes during this process. Total RNAs were extracted from human induced pluripotent stem cells and induced pluripotent stem cell-derived neural progenitor cells. Their gene expression profiles were investigated using the Affymetrix GeneChip Human Transcriptome Array 2.0 platform.
Project description:RNA (poly(A)+ fraction) has ben isolated from undifferentiated mouse embryonic stem cells and from embryonic stem cells induced to differentiate using the hanging drop model, 6 days following iinduction of differentiation. Profiling of the transcritome at these two stages of differentiation using deep RNA sequencing allows identifying modulated coding and noncoding transcripts upon induction of differentiation. RNA profiling of mouse embryonic stem cells at two stages of differentiation, i.e. undifferntiated state and at day 6 after induction of differentiation. Two conditions, three biological replicates per condition
Project description:Pnpla6 gene was silenced in D3 mouse embryonic stem cells and the cells were allowed to spontaneously differentiate. Transcription of the whole genome was analyzed 96 days afterwards and compared against control cells differentiated during the same time without previous silencing.
