E-GEOD-46854 - Induction of the mouse germ cell fate by transcription factors in vitro [exp2]

Status
Released on 4 August 2013, last updated on 3 June 2014
Organism
Mus musculus
Samples (20)
Array (1)
Protocols (7)
Description
The germ cell lineage ensures the continuity of life through the generation of male and female gametes, which unite to form a totipotent zygote. We have established a culture system that recapitulates the mouse germ-cell specification pathway: Using cytokines, embryonic stem cells (ESCs)/induced pluripotent stem cells (iPSCs) are induced into epiblast-like cells (EpiLCs) and then into primordial germ cell-like cells (PGCLCs) with capacity both for spermatogenesis and oogenesis, creating an opportunity for understanding and regulating mammalian germ cell development in both sexes in vitro. Here we show that, without cytokines, simultaneous over-expression of three transcription factors (TFs), Blimp1 (also known as Prdm1), Prdm14 and Tfap2c (also known as AP2γ), directs EpiLCs, but not ESCs, swiftly and highly efficiently into a PGC state with endogenous transcription circuitry. The induction of the PGC state on EpiLCs minimally requires Prdm14 but not Blimp1 or Tfap2c. The TF-induced PGC state reconstitutes key transcriptome and epigenetic reprogramming in PGCs, but bypasses a mesodermal program that accompanies PGC specification in vivo and in vitro by cytokines including BMP4. Importantly, the TF-induced PGC-like cells robustly contribute to spermatogenesis and fertile offspring. Our findings provide not only a novel insight into the transcriptional logic that creates a germ cell state, but also a foundation for the TF-based reconstitution and regulation of mammalian gametogenesis. Aim of this analysis is identification of genes whose expression was altered by each of key transcription factor for transcription factor-induced primordial germ cells (TF-PGCLCs) induction (Blimp1 (B), Prdm14 (P14), and Tfap2c (A)). Both of epiblast-like cells (EpiLCs) (Hayashi et al., 2011, Cell) and aggregates of EpiLCs cultured with doxycycline on day 1 were harvested for 5 cell lines, including BP14A (Clone #3-3), B (#2-4), P14 (#7-109), A (#8-2), and the parental clone (BVSCR26rtTA embryonic stem cells). Total RNA was isolated and analyzed. Two biological duplicates for each cell type were included.
Experiment type
transcription profiling by array 
Contacts
Fumio NAKAKI <nakaki@anat2.med.kyoto-u.ac.jp>, Fumio Nakaki, Hiroshi Ohta, Katsuhiko Hayashi, Kazuki Kurimoto, Mitinori Saitou, Yukihiro Yabuta
Citation
Induction of mouse germ-cell fate by transcription factors in vitro. Nakaki F, Hayashi K, Ohta H, Kurimoto K, Yabuta Y, Saitou M. , Europe PMC 23913270
MIAME
PlatformsProtocolsFactorsProcessedRaw
Files
Investigation descriptionE-GEOD-46854.idf.txt
Sample and data relationshipE-GEOD-46854.sdrf.txt
Raw data (1)E-GEOD-46854.raw.1.zip
Processed data (1)E-GEOD-46854.processed.1.zip
Array designA-AFFY-45.adf.txt
Links