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RNA sequencing of conventional and reset human pluripotent stem cells

ABSTRACT: Human pluripotent cells were reset to ground state pluripotency by transient overexpression of NANOG and KLF2 and subsequent inhibition of ERK and protein kinase C. Transcriptional profiling of reset cells and conventional pluripotent stem cell cultures was carried out by RNA-seq, in tandem with mouse embryonic stem cells propagated under similar conditions to assess the combinatorial effects of MEK inhibitor PD0325901, GSK3 inhibitor CHIR99021 and PKC inhibitor Go6983.

INSTRUMENT(S): Illumina HiSeq 2000

ORGANISM(S): Mus musculus

SUBMITTER: Paul Bertone

PROVIDER: E-MTAB-2857 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Resetting transcription factor control circuitry toward ground-state pluripotency in human.

Takashima Yasuhiro Y Guo Ge G Loos Remco R Loos Remco R Nichols Jennifer J Ficz Gabriella G Krueger Felix F Oxley David D Santos Fatima F Clarke James J Mansfield William W Reik Wolf W Bertone Paul P Smith Austin A

Cell 20140901 6

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 sustains a transgene-independent rewired state. Reset cells self-renew continuously without ERK signaling, are phenotypically stable, and are kary ...[more]

PMID: 25215486

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